加减逍遥散影响慢性心理应激T2DM小鼠IR机制的研究
摘要
目的：
糖尿病是当今社会发病率仅次于心脑血管及癌症的第三类重大身心疾病。我国糖尿病发病率高达9.7%,糖尿病病人接近一个亿,中国成为全球“糖尿病第一大国”。糖尿病中90%以上为T2DM, T2DM及其相关并发症已成为威胁人类身心健康的重大问题。从糖尿病的病因研究来看,现代新的医学模式生物---心理---社会医学模式认为,糖尿病是一种心理和身体相关的心身疾病,心理应激是糖尿病的一个重要的诱发因素,而且社会心理因素越来越为人们重视,而这与中医理论中的“情志致病”理论相合。
T2DM中80%以上存在胰岛素抵抗(IR)。IR是T2DM的主要发病机制之一,伴随着T2DM发生、发展的全过程,同时也是肥胖、高血压和血脂异常等多种代谢疾病的重要病理基础。但引起IR的病理机制十分复杂且目前尚不十分清楚。因此研究IR,特别是从中西医结合的方向来研究IR的发生机制,不仅能够丰富中西医的理论,还能对各种代谢性疾病有一定的预防和治疗的作用。
本研究根据糖尿病的临床特点以及中医有关“情志病因最先伤肝”的理论,提出糖尿病的基本病机是脾失健运、不能化精,情志内伤,木失疏泄,土虚木乘是糖尿病发展的重要诱因的假说。以心理应激自发性糖尿病小鼠为研究对象,采用病理学、分子生物学等现代实验技术,从糖代谢及相关基因变化探讨心理应激影响T2DM小鼠IR的机理及中药复方加减逍遥散的作用机制,为中医药防治糖尿病探索新的途径。
方法：
以自发性糖尿病雄性小鼠,高糖高脂饲料喂养连续8周以复制T2DM小鼠模型,通过电击、夹尾、禁食、禁水、合笼饲养等方法复制慢性心理应激模型。观察加味逍遥散对慢性心理应激T2DM小鼠血液生化指标的影响；加味逍遥散对慢性心理应激T2DM小鼠相关基因表达的影响；加味逍遥散对慢性心理应激T2DM小鼠GR表达的影响以及加味逍遥散对慢性心理应激T2DM小鼠IRS-1表达的影响。
结果：
1加味逍遥散对慢性心理应激T2DM小鼠血液生化指标的影响
与空白对照组(WC组)比较,应激组(SC组)的空腹血糖(FBG)有所升高，而糖化血红蛋白(HbAlc)明显升高,胰岛素敏感性指数(ISI)明显降低,体重明显下降,甘油三酯(TG)、总胆固醇(TCHO)明显升高,高密度脂蛋白胆固醇(HDL-C)明显降低,XYS组TG、CHO、LDL-C降低,HDL-C显著升高；与SC组比较,应激加逍遥散低剂量组(S+LXYS组)及应激加逍遥散高剂量组(S+HXYS组)的FBG有下降的趋势,而HbAlc则明显下降,S+HXYS组体重明显升高,TG, CHO, LDL-C明显降低,HDL-C明显升高。
2加味逍遥散对慢性心理应激T2DM小鼠相关基因表达的影响
在89个相关基因中,与WC组相比较,SC组表达上调的基因有FOXC2、Ifng、IL-10、 Sell、lc2a4、Tnf及Ucp2等7个,下调的基因有Aqp2、cr2、FOXgl、FOXP3、fkfb3、 Tnfrsflb等6个；XYS组上调的基因有Adral-α、FOXC2、Ifng、Neurod1、Nsf. Pparg. Slc2a4、Gusb等8个,下调的基因有Aqp2、Pfkfb3、Tnfrsflb等3个。与SC组相比较,S+LXYS组上调的基因FOX P3、Ppargcla等2个,下调的基因有Ace. IL-10. Slc2a4. Aktb等4个；S+HXYS组上调的基因有Acly. Adral-α、Aqp2、PP4、Fbp1、G6PC、Gpd1、 Icam1、IgfbpS. IL-4ra等37个,下调的基因有Adra3-β、GIPlr、Vegfa、Actb、Snap25、Pax4等6个。
3加味逍遥散对慢性心理应激T2DM小鼠GR表达的影响
3.1各组小鼠胰岛细胞内GR表达差异
从形态学特征分析,SC组胰岛部位荧光强度最强,且与周围胰腺组织边界清晰；其次是WC组,但其荧光强度与周围胰腺组织反差较小；再其次为S+LXYS组及S+HXYS组；荧光强度最小的是XYS组。从统计数据分析：与WC组相比较,SC组小鼠胰岛细胞内GR表达明显上升,XYS组则明显下降；与SC组相比较,S+LXYS组与S+HXYS组胰岛细胞内GR均明显降低。
3.2各组小鼠肝细胞内GR表达差异
从形态学特征分析,各组小鼠肝细胞GR表达所显示出的荧光强度依次为WC组、S+HXYS组、XYS组、S+LXYS组及SC组。各组小鼠肝组织的微观结构存在较大差异,WC组肝细胞结构完整,但排列不太规则,放射状肝索结构不明显,细胞体相对较大,XYS组肝组织细胞排列比较规则,可见明显的放射状肝索结构,而SC组的肝细胞排列较乱而稀疏,并有许多空泡状结构；S+LXYS组与S+HXYS组在肝细胞数目与排列上均有所恢复,但以S+HXYS组更明显。从统计数据结果分析：与WC组相比较,SC组与XYS组小鼠肝细胞内GR表达明显下降；与SC组相比较,S+LXYS组肝细胞内GR有所上升。而S+HXYS组则明显上升,但S+LXYS组及S+HXYS组均明显低于WC组。
4加味逍遥散对慢性心理应激T2DM小鼠工RS-1表达的影响
4.1各组小鼠胰岛细胞IRS-1表达比较
经DAPI复染后,荧光显微镜下可以清晰分辨各组小鼠胰岛细胞分布,WC组与SC组小鼠胰岛细胞分布相对稀疏,绿色荧光较弱,XYS组、S+LXYS组及S+HXYS组荧光强度较高。从统计数据结果分析也进一步证实：与WC组相比较,SC组IRS-1表达下调,XYS组IRS-1表达上调：与SC组相比较,S+LXYS组与S+HXYS组IRS1表达上调。
4.2各组小鼠肝细胞IRS-1表达比较
从各组小鼠肝细胞免疫荧光及DAPI复染图片比较发现：WC组及SC组小鼠肝细胞排列较乱而稀疏,肝索结构不清晰,细胞核也大小不一,可能是肝细胞死亡后代偿性再生出一些新的细胞,绿色荧光较弱：XYS组及S+HXYS组肝细胞排列较整齐,肝素结构比较清晰,绿色荧光较强,S+LXYS组则介于二者之间。从统计数据也进一步显示：与WC组相比较,SC组肝细胞内绿色荧光强度降低,XYS组的绿色荧光强度增大；与SC组相比较,S+LXYS组绿色荧光稍有增强,而S+HXYS组绿色荧光显著增强。
结论：
1慢性心理应激能加重T2DM小鼠的高血糖病情,降低其胰岛素的敏感性,升高其血清中的TG、CHO,降低HDL-C,使小鼠的体重相对减轻；加味逍遥散一方面具有降糖作用,对T2DM具有较好的治疗效果,另一方面也能对抗慢性心理应激所致的血糖升高,增加胰岛素的敏感性,降低T2DM小鼠血清中的TG、CHO、LDL-C,升高HDL-C,并且减轻应激对小鼠血脂及体重的影响。其作用机制可能是通过提高胰岛素的敏感性从而纠正了T2DM小鼠糖脂代谢的紊乱。
2由于T2DM遗传背景的多样性及异质性,目前对多基因遗传性疾病,尚缺乏正确、全面、合适的技术策略以及家系遗传和疾病的连锁分析法。本实验选择了89个T2DM相关基因,采用PCR芯片技术研究探讨慢性心理应激对T2DM小鼠IR的影响及加减逍遥散的作用机制。结果发现：与WC组相比较,SC组上调的基因有7个,下调的基因有6个；XYS组上调的基因有8个,下调有3个。SC组与XYS组均上调的基因有FOXC2、Ifng、Slc2a4等三个,均下调的基因有下调的基因有Aqp2、Pfkfb3、Tnfrsflb等3个。因此,慢性心理应激可能是通过调节T2DM多种相关基因表达,加重了IR;而加减逍遥散能够对T2DM具有防治作用以及改善IR,也与其调节T2DM相关基因表达密切相关。
3慢性心理应激可使T2DM小鼠胰岛细胞内的GR表达上调,而加减逍遥散则可降低T2DM小鼠胰岛细胞内GR的表达,对于慢性应激模型下的T2DM小鼠而言,S+LXYS组与S+HXYS组均能降低其GR在胰岛细胞中的表达,且S+HXYS组的效果优于S+LXYS组。但对于肝细胞而言,SC组的GR表达反而下降,这可能是与长期慢性应激导致部分肝细胞坏死有关,S+LXYS组与S+HXYS组的GR表达有所回升,且S+HXYS组上升更明显,说明加味逍遥散能对抗慢性心理应激对肝细胞的损伤。
4慢性心理应激能导致T2DM小鼠胰岛细胞及肝细胞内IRS-1表达下调,而加减逍遥散能上调T2DM小鼠胰岛细胞及肝细胞内IRS-1表达,这与第二节的实验中IRS-1基因表达RT-PCR(?)勺结果相一致。说明抑制IRS-1在胰岛β细胞的表达,减少胰岛素的分泌，以及抑制IRS-1在肝细胞的表达,降低肝糖元的合成可能是慢性心理应激加重T2DM小鼠IR的重要机制,而加减逍遥散则可能是通过促进胰岛β细胞及肝细胞内IRS-1的表达,促进胰岛素的分泌及其生物利用度,加快肝糖元的合成而发挥降低血糖,缓解IR的功效。
Objective
Diabetes is the third important mental and physical disease preceded only by thd heart-brain artery and cancer. It's incidence surpass9.7%. T he disease number of people reach about100million. Now China is the max country of diabetes people in the world. Among exceeding90%is Type2diabetes. T2DM and it's complications have becomed the serious psychologi cal-physical problem to threaten the humanity. The new medicical mode of organism-psychology-society consider that the diabetes is a kind of the p sychological and physical disease. Psychological sensitive is the importa nt inducing factor. And the social and psychological factors have given t o value by the people, which is indentical with "feeling disese" of the theory of the traditional Chinese medicine.
The occurrence of insulin resistance is surpass80%in T2DM. IR is o ne kind of the primary disease mechanism. It appears the whole process of the happening and development in the T2DM, At the same time, it is also the important pathological foundation of many metabolic diseases such as obesity, hypertension and hyperlipdimia and so on. But now, IR's patholog ical mechanism is very complex and vague. So reseaching IR, especially st udying it's occurrence mechanism from the direction of combining traditio nal Chinese and Western medicine, which it is not only enrich the theory of traditional Chinese and Western medicine, but also can prevent and cur e many metabolic diseases.
The study put forward to a important hypothesis, that the diabetes's basic phthogenesis is the spleen can not generate the essence, according t o the diabetes clinical characteristic and feeling disease theory. The st udy object is spontaneous diabetes mouse, method is pathology, molecular biology, and from the sugar-substitution and gene to probe the the mechan ism of the mental sensitive influence IR and the effect mechanism of the compound prescription of Xiao-yao power medicine.
Methods
The study uses male KK mouses, feeding the fodder of hyperglycemia-hy perlipidemia for8weeks to build the model of T2DM. Using electric-shock, clamping the tail,not eating, not water and so on, to build the model of ch ronic-psychological-stress. The purposes to observe the Xiao-Yao-San's eff ect to biochemistry index of chronic—mental-irritable T2DM mouses, the e ffect to gene express of chronic-psychological-stress T2DM mouses, the ef feet to glucocorticosteroid receptor(GR) express and IRS-1express of chr onic-psychological-stress T2DM mouses.
Results
1The Xiao-Yao-San's effect to biochemistry index of chronic--psychol ogical-stress T2DM mouses
Compare to WC-group, SC-group mouses'FBG raise, HbAlc ascend and ISI descend clearly. Their's body weight reduce, TG、CHO ascend and HDL-C desc end. But the XYS-group are opposited. To compare SC-group, the groups'of S+LXYS and S+HXYS FBG and HbAlc descend, the body weight ascend, TG,CHO,L DL-C descend and HDL-C ascend clearly.
2The Xiao-Yao-San's effect to gene express of chronic--psychological-stress T2DM mouses
Among89related genes, comparing to WC-group, there7genes ascend, FO XC2、Ifng、IL-10、Sell、Slc2a4、Tnf and Ucp2and6genes descend, Aqp2, Cc r2、FOXg1、F0XP3、Pfkfb3、Tnfrsflb in SC-group. There are8genes ascend, A dral-α、FOXC2、Ifng、Neurodl、Nsf、Pparg、Slc2a4、Gusb and3genes descen d, Aqp2、Pfkfb3、Tnfrsf1in XYS-group. Comparing to SC-group,there are2g enes ascend, FOX P3、Ppargcla,4genes descend, Ace、IL-10、Slc2a4, Aktb in S+LXYS-group. There are37genes ascend, Ac1y、Adral-α、Aqp2、DPP4、Fbp1、 G6PC、Gpd1、Icaml、Igfbp5、IL-4ra、Insl、Mapkl4、Nfkb1、Nos3、Nrf1、Nsf、P arpl、Pik3cd、Ppara、Pparg、Ptpn1、Pyg1、Rab4a、Slcl4a2、Snap23、Sod2、Sre bf1、Stxbp1、Tnfrsfla、Trib3、Ucp2、Vamp3、Yapa、Hprt、Hsp90abl,6descend genes, Adra3-β、GlPlr、Vegfa、Actb、Snap25、Pax4.
3The Xiao-Yao-san's effect to GR express of chronic-psychological-s tress T2DM mouses
3.1the GR express difference in insulin cells of all groups
Analysis from morphology, SC-group's fluorescence strength is the bes t intheinsulin, and the better is WC-group, and in order is S+LXYS and S+HXYS groups, the weakest is XYS-group. Analysis from statistical data, th e results are the same, that is comparing WC-group, SC-group's GR express in insulin cells ascend clearly, and XYS-group is opposite results. Comp are SC-group, GR express in insulin of S+LXYS and S+HXYS groups are desce nd.
3.2the GR express difference in liver cells of all groups
Analysis from morphology,the fluorescence strength is followed from s trong to weak, WC-group, S+HXYS group,XYS-group, S+LXYS-group and SC-group. The micro-structures are different. The liver-cell structures are intact of WC-group, but the array are not rule and the radioactive liver-rope st ructure are fuzzy. The liver-cell array are rule and radioactive liver-ro pe structure are clear of XYS-group. But the liver-cell array are disorde r and infrequent and have many bubbles. The liver cell array and numbers are better than SC-group. Analysis from statistical data, comparing WC-gr oup, GR express in liver cells ascend clearly of SC and XYS group. Compar ing SC-group, GR express in liver cells of S+LXYS and S+HXYS groups are a scend clearly. But the two groups are lower than WC-group.
4The Xiao-Yao-San's effect to IRS-1express of chronic-psychologica1-stress T2DM mouses
4.1the IRS-1express difference in insulin cells of all groups
Analysis from morphology, the green fluorescence strength of XYS, S+LX YS and groups are clearer than WC and SC groups. Analysis from statistica1data, comparing WC-group, IRS-1express in insulin cells descend and as cend of SC group. Comparing SC-group, IRS-1express in insulin cells of a re ascend of S+LXYS and S+HXYS groups.
4.2the IRS-1express difference in liver cells of all groups
Analysis from morphology, the green fluorescence strength of S+HXYS,X YS, and S+LXYS groups are clear and the liver array are rule and liver ro pe clear,but the green fluorescence strength of WC and SC groups are weak and the liver cells array disorder and liver rope structures fuzzy. Anal 'sis from statistical data, comparing WC-group, IRS-1express in liver ce lls descend of SC-group and ascend of XYS-group. Comparing SC-group, IRS- 1express in liver cells are ascend of S+LXYS and S+HXYS groups, between the S+HXYS group is clearer.
conelusions
1Chronic-psychological-stress can make more serious hyperglycemia f or T2DM, reduce insuline sensitivity, raise TG、CHO,reduce the body weight. XYS-power'effect lies in the following:one aspect can descend the gluco se,the second can resist the hyperglycemia resulted from Chronic-psycholo gical-stress, ascend insuline sensitivity, reduce TG、CHO、LDL-C, ascend HDL-C, and reduce hyperlipidemia and body weight becaused of Chronic-psychol ogical-stress. It's effect mechanism might have increased insuline sensit ivity to correct the sugar-fat disorder.
2Because of the variegation and heterogeneity of T2DM heredity, the re is not proper, comprehensive, suitable technology and heredity analysis methods for gene-heredity diseases. The study selects89related gene of T2DM, uses PCR microchip technology to probe the influence of Chronic-psyc hological-stress to IR of T2DM and Xiao-Yao-San effect machnism. Result:c omparing to WC-group,there is7genes ascending,6genes descending of SC-group; there is8genes ascending,3genes descending of XYS-group; among FOXC2、Ifng、Slc2a4are their common ascending genes. Aqp2、Pfkfb3. Tnfrsf1b are their common descending genes.So, Chronic-psychological-stress can adjust related genes express and make more serious IR of T2DM. Xiao-Yao-San can prevent and cure diabetes and improve IR, Which it is might relate d to genes express.
3Chronic-psychological-stress can increase GR express in insulin ce11of T2DM mouses.But Xiao-Yao-San can reduce GR express in the insulin c ell of T2DM. S+LXYS and S+HXYS groups all can reduce GR express in insulin cell of Chronic-psychological-stress T2DM, and S+HDL-C group's effect is better than S+LXYS group. But GR express in liver cell of SC-group is desc end, that can relate to liver cell death resulted from long-time Chronic-p sychological-stress.GR express in liver cell of S+LXYS and S+HXYS groups are ascend, and S+HXYS group's effect is better than S+LXYS group, which it might be Xiao-Yao-San make better liver cell damage becaused of Chronic-psychol ogical-stress.
4Chronic-psychological-stress can decrease IRS-1express in insulin cell of T2DM mouses.But Xiao-Yao-San ascend IRS-1express in insulin cel l and liver cell of T2DM mouses, that the result is the same to the gene e xpress. The result is to show that inhibiting IRS-1express in insulin ce ll,reducing insulin secretion, and inhibiting IRS-1express in liver cell, reducing liver glycogen's compound might be important effect mechanism of IR of T2DM mouses resulted from Chronic-psychological-stress. Xiao-Yao-S an can ascend IRS-1express in insulin and liver cells, increase insulin s ectetion, increase liver glycogen's compound to reduce glucose and relieve insulin resistant.
糖尿病是当今社会发病率仅次于心脑血管及癌症的第三类重大身心疾病。我国糖尿病发病率高达9.7%,糖尿病病人接近一个亿,中国成为全球“糖尿病第一大国”。糖尿病中90%以上为T2DM, T2DM及其相关并发症已成为威胁人类身心健康的重大问题。从糖尿病的病因研究来看,现代新的医学模式生物---心理---社会医学模式认为,糖尿病是一种心理和身体相关的心身疾病,心理应激是糖尿病的一个重要的诱发因素,而且社会心理因素越来越为人们重视,而这与中医理论中的“情志致病”理论相合。
T2DM中80%以上存在胰岛素抵抗(IR)。IR是T2DM的主要发病机制之一,伴随着T2DM发生、发展的全过程,同时也是肥胖、高血压和血脂异常等多种代谢疾病的重要病理基础。但引起IR的病理机制十分复杂且目前尚不十分清楚。因此研究IR,特别是从中西医结合的方向来研究IR的发生机制,不仅能够丰富中西医的理论,还能对各种代谢性疾病有一定的预防和治疗的作用。
本研究根据糖尿病的临床特点以及中医有关“情志病因最先伤肝”的理论,提出糖尿病的基本病机是脾失健运、不能化精,情志内伤,木失疏泄,土虚木乘是糖尿病发展的重要诱因的假说。以心理应激自发性糖尿病小鼠为研究对象,采用病理学、分子生物学等现代实验技术,从糖代谢及相关基因变化探讨心理应激影响T2DM小鼠IR的机理及中药复方加减逍遥散的作用机制,为中医药防治糖尿病探索新的途径。
方法：
以自发性糖尿病雄性小鼠,高糖高脂饲料喂养连续8周以复制T2DM小鼠模型,通过电击、夹尾、禁食、禁水、合笼饲养等方法复制慢性心理应激模型。观察加味逍遥散对慢性心理应激T2DM小鼠血液生化指标的影响；加味逍遥散对慢性心理应激T2DM小鼠相关基因表达的影响；加味逍遥散对慢性心理应激T2DM小鼠GR表达的影响以及加味逍遥散对慢性心理应激T2DM小鼠IRS-1表达的影响。
结果：
1加味逍遥散对慢性心理应激T2DM小鼠血液生化指标的影响
与空白对照组(WC组)比较,应激组(SC组)的空腹血糖(FBG)有所升高，而糖化血红蛋白(HbAlc)明显升高,胰岛素敏感性指数(ISI)明显降低,体重明显下降,甘油三酯(TG)、总胆固醇(TCHO)明显升高,高密度脂蛋白胆固醇(HDL-C)明显降低,XYS组TG、CHO、LDL-C降低,HDL-C显著升高；与SC组比较,应激加逍遥散低剂量组(S+LXYS组)及应激加逍遥散高剂量组(S+HXYS组)的FBG有下降的趋势,而HbAlc则明显下降,S+HXYS组体重明显升高,TG, CHO, LDL-C明显降低,HDL-C明显升高。
2加味逍遥散对慢性心理应激T2DM小鼠相关基因表达的影响
在89个相关基因中,与WC组相比较,SC组表达上调的基因有FOXC2、Ifng、IL-10、 Sell、lc2a4、Tnf及Ucp2等7个,下调的基因有Aqp2、cr2、FOXgl、FOXP3、fkfb3、 Tnfrsflb等6个；XYS组上调的基因有Adral-α、FOXC2、Ifng、Neurod1、Nsf. Pparg. Slc2a4、Gusb等8个,下调的基因有Aqp2、Pfkfb3、Tnfrsflb等3个。与SC组相比较,S+LXYS组上调的基因FOX P3、Ppargcla等2个,下调的基因有Ace. IL-10. Slc2a4. Aktb等4个；S+HXYS组上调的基因有Acly. Adral-α、Aqp2、PP4、Fbp1、G6PC、Gpd1、 Icam1、IgfbpS. IL-4ra等37个,下调的基因有Adra3-β、GIPlr、Vegfa、Actb、Snap25、Pax4等6个。
3加味逍遥散对慢性心理应激T2DM小鼠GR表达的影响
3.1各组小鼠胰岛细胞内GR表达差异
从形态学特征分析,SC组胰岛部位荧光强度最强,且与周围胰腺组织边界清晰；其次是WC组,但其荧光强度与周围胰腺组织反差较小；再其次为S+LXYS组及S+HXYS组；荧光强度最小的是XYS组。从统计数据分析：与WC组相比较,SC组小鼠胰岛细胞内GR表达明显上升,XYS组则明显下降；与SC组相比较,S+LXYS组与S+HXYS组胰岛细胞内GR均明显降低。
3.2各组小鼠肝细胞内GR表达差异
从形态学特征分析,各组小鼠肝细胞GR表达所显示出的荧光强度依次为WC组、S+HXYS组、XYS组、S+LXYS组及SC组。各组小鼠肝组织的微观结构存在较大差异,WC组肝细胞结构完整,但排列不太规则,放射状肝索结构不明显,细胞体相对较大,XYS组肝组织细胞排列比较规则,可见明显的放射状肝索结构,而SC组的肝细胞排列较乱而稀疏,并有许多空泡状结构；S+LXYS组与S+HXYS组在肝细胞数目与排列上均有所恢复,但以S+HXYS组更明显。从统计数据结果分析：与WC组相比较,SC组与XYS组小鼠肝细胞内GR表达明显下降；与SC组相比较,S+LXYS组肝细胞内GR有所上升。而S+HXYS组则明显上升,但S+LXYS组及S+HXYS组均明显低于WC组。
4加味逍遥散对慢性心理应激T2DM小鼠工RS-1表达的影响
4.1各组小鼠胰岛细胞IRS-1表达比较
经DAPI复染后,荧光显微镜下可以清晰分辨各组小鼠胰岛细胞分布,WC组与SC组小鼠胰岛细胞分布相对稀疏,绿色荧光较弱,XYS组、S+LXYS组及S+HXYS组荧光强度较高。从统计数据结果分析也进一步证实：与WC组相比较,SC组IRS-1表达下调,XYS组IRS-1表达上调：与SC组相比较,S+LXYS组与S+HXYS组IRS1表达上调。
4.2各组小鼠肝细胞IRS-1表达比较
从各组小鼠肝细胞免疫荧光及DAPI复染图片比较发现：WC组及SC组小鼠肝细胞排列较乱而稀疏,肝索结构不清晰,细胞核也大小不一,可能是肝细胞死亡后代偿性再生出一些新的细胞,绿色荧光较弱：XYS组及S+HXYS组肝细胞排列较整齐,肝素结构比较清晰,绿色荧光较强,S+LXYS组则介于二者之间。从统计数据也进一步显示：与WC组相比较,SC组肝细胞内绿色荧光强度降低,XYS组的绿色荧光强度增大；与SC组相比较,S+LXYS组绿色荧光稍有增强,而S+HXYS组绿色荧光显著增强。
结论：
1慢性心理应激能加重T2DM小鼠的高血糖病情,降低其胰岛素的敏感性,升高其血清中的TG、CHO,降低HDL-C,使小鼠的体重相对减轻；加味逍遥散一方面具有降糖作用,对T2DM具有较好的治疗效果,另一方面也能对抗慢性心理应激所致的血糖升高,增加胰岛素的敏感性,降低T2DM小鼠血清中的TG、CHO、LDL-C,升高HDL-C,并且减轻应激对小鼠血脂及体重的影响。其作用机制可能是通过提高胰岛素的敏感性从而纠正了T2DM小鼠糖脂代谢的紊乱。
2由于T2DM遗传背景的多样性及异质性,目前对多基因遗传性疾病,尚缺乏正确、全面、合适的技术策略以及家系遗传和疾病的连锁分析法。本实验选择了89个T2DM相关基因,采用PCR芯片技术研究探讨慢性心理应激对T2DM小鼠IR的影响及加减逍遥散的作用机制。结果发现：与WC组相比较,SC组上调的基因有7个,下调的基因有6个；XYS组上调的基因有8个,下调有3个。SC组与XYS组均上调的基因有FOXC2、Ifng、Slc2a4等三个,均下调的基因有下调的基因有Aqp2、Pfkfb3、Tnfrsflb等3个。因此,慢性心理应激可能是通过调节T2DM多种相关基因表达,加重了IR;而加减逍遥散能够对T2DM具有防治作用以及改善IR,也与其调节T2DM相关基因表达密切相关。
3慢性心理应激可使T2DM小鼠胰岛细胞内的GR表达上调,而加减逍遥散则可降低T2DM小鼠胰岛细胞内GR的表达,对于慢性应激模型下的T2DM小鼠而言,S+LXYS组与S+HXYS组均能降低其GR在胰岛细胞中的表达,且S+HXYS组的效果优于S+LXYS组。但对于肝细胞而言,SC组的GR表达反而下降,这可能是与长期慢性应激导致部分肝细胞坏死有关,S+LXYS组与S+HXYS组的GR表达有所回升,且S+HXYS组上升更明显,说明加味逍遥散能对抗慢性心理应激对肝细胞的损伤。
4慢性心理应激能导致T2DM小鼠胰岛细胞及肝细胞内IRS-1表达下调,而加减逍遥散能上调T2DM小鼠胰岛细胞及肝细胞内IRS-1表达,这与第二节的实验中IRS-1基因表达RT-PCR(?)勺结果相一致。说明抑制IRS-1在胰岛β细胞的表达,减少胰岛素的分泌，以及抑制IRS-1在肝细胞的表达,降低肝糖元的合成可能是慢性心理应激加重T2DM小鼠IR的重要机制,而加减逍遥散则可能是通过促进胰岛β细胞及肝细胞内IRS-1的表达,促进胰岛素的分泌及其生物利用度,加快肝糖元的合成而发挥降低血糖,缓解IR的功效。
Objective
Diabetes is the third important mental and physical disease preceded only by thd heart-brain artery and cancer. It's incidence surpass9.7%. T he disease number of people reach about100million. Now China is the max country of diabetes people in the world. Among exceeding90%is Type2diabetes. T2DM and it's complications have becomed the serious psychologi cal-physical problem to threaten the humanity. The new medicical mode of organism-psychology-society consider that the diabetes is a kind of the p sychological and physical disease. Psychological sensitive is the importa nt inducing factor. And the social and psychological factors have given t o value by the people, which is indentical with "feeling disese" of the theory of the traditional Chinese medicine.
The occurrence of insulin resistance is surpass80%in T2DM. IR is o ne kind of the primary disease mechanism. It appears the whole process of the happening and development in the T2DM, At the same time, it is also the important pathological foundation of many metabolic diseases such as obesity, hypertension and hyperlipdimia and so on. But now, IR's patholog ical mechanism is very complex and vague. So reseaching IR, especially st udying it's occurrence mechanism from the direction of combining traditio nal Chinese and Western medicine, which it is not only enrich the theory of traditional Chinese and Western medicine, but also can prevent and cur e many metabolic diseases.
The study put forward to a important hypothesis, that the diabetes's basic phthogenesis is the spleen can not generate the essence, according t o the diabetes clinical characteristic and feeling disease theory. The st udy object is spontaneous diabetes mouse, method is pathology, molecular biology, and from the sugar-substitution and gene to probe the the mechan ism of the mental sensitive influence IR and the effect mechanism of the compound prescription of Xiao-yao power medicine.
Methods
The study uses male KK mouses, feeding the fodder of hyperglycemia-hy perlipidemia for8weeks to build the model of T2DM. Using electric-shock, clamping the tail,not eating, not water and so on, to build the model of ch ronic-psychological-stress. The purposes to observe the Xiao-Yao-San's eff ect to biochemistry index of chronic—mental-irritable T2DM mouses, the e ffect to gene express of chronic-psychological-stress T2DM mouses, the ef feet to glucocorticosteroid receptor(GR) express and IRS-1express of chr onic-psychological-stress T2DM mouses.
Results
1The Xiao-Yao-San's effect to biochemistry index of chronic--psychol ogical-stress T2DM mouses
Compare to WC-group, SC-group mouses'FBG raise, HbAlc ascend and ISI descend clearly. Their's body weight reduce, TG、CHO ascend and HDL-C desc end. But the XYS-group are opposited. To compare SC-group, the groups'of S+LXYS and S+HXYS FBG and HbAlc descend, the body weight ascend, TG,CHO,L DL-C descend and HDL-C ascend clearly.
2The Xiao-Yao-San's effect to gene express of chronic--psychological-stress T2DM mouses
Among89related genes, comparing to WC-group, there7genes ascend, FO XC2、Ifng、IL-10、Sell、Slc2a4、Tnf and Ucp2and6genes descend, Aqp2, Cc r2、FOXg1、F0XP3、Pfkfb3、Tnfrsflb in SC-group. There are8genes ascend, A dral-α、FOXC2、Ifng、Neurodl、Nsf、Pparg、Slc2a4、Gusb and3genes descen d, Aqp2、Pfkfb3、Tnfrsf1in XYS-group. Comparing to SC-group,there are2g enes ascend, FOX P3、Ppargcla,4genes descend, Ace、IL-10、Slc2a4, Aktb in S+LXYS-group. There are37genes ascend, Ac1y、Adral-α、Aqp2、DPP4、Fbp1、 G6PC、Gpd1、Icaml、Igfbp5、IL-4ra、Insl、Mapkl4、Nfkb1、Nos3、Nrf1、Nsf、P arpl、Pik3cd、Ppara、Pparg、Ptpn1、Pyg1、Rab4a、Slcl4a2、Snap23、Sod2、Sre bf1、Stxbp1、Tnfrsfla、Trib3、Ucp2、Vamp3、Yapa、Hprt、Hsp90abl,6descend genes, Adra3-β、GlPlr、Vegfa、Actb、Snap25、Pax4.
3The Xiao-Yao-san's effect to GR express of chronic-psychological-s tress T2DM mouses
3.1the GR express difference in insulin cells of all groups
Analysis from morphology, SC-group's fluorescence strength is the bes t intheinsulin, and the better is WC-group, and in order is S+LXYS and S+HXYS groups, the weakest is XYS-group. Analysis from statistical data, th e results are the same, that is comparing WC-group, SC-group's GR express in insulin cells ascend clearly, and XYS-group is opposite results. Comp are SC-group, GR express in insulin of S+LXYS and S+HXYS groups are desce nd.
3.2the GR express difference in liver cells of all groups
Analysis from morphology,the fluorescence strength is followed from s trong to weak, WC-group, S+HXYS group,XYS-group, S+LXYS-group and SC-group. The micro-structures are different. The liver-cell structures are intact of WC-group, but the array are not rule and the radioactive liver-rope st ructure are fuzzy. The liver-cell array are rule and radioactive liver-ro pe structure are clear of XYS-group. But the liver-cell array are disorde r and infrequent and have many bubbles. The liver cell array and numbers are better than SC-group. Analysis from statistical data, comparing WC-gr oup, GR express in liver cells ascend clearly of SC and XYS group. Compar ing SC-group, GR express in liver cells of S+LXYS and S+HXYS groups are a scend clearly. But the two groups are lower than WC-group.
4The Xiao-Yao-San's effect to IRS-1express of chronic-psychologica1-stress T2DM mouses
4.1the IRS-1express difference in insulin cells of all groups
Analysis from morphology, the green fluorescence strength of XYS, S+LX YS and groups are clearer than WC and SC groups. Analysis from statistica1data, comparing WC-group, IRS-1express in insulin cells descend and as cend of SC group. Comparing SC-group, IRS-1express in insulin cells of a re ascend of S+LXYS and S+HXYS groups.
4.2the IRS-1express difference in liver cells of all groups
Analysis from morphology, the green fluorescence strength of S+HXYS,X YS, and S+LXYS groups are clear and the liver array are rule and liver ro pe clear,but the green fluorescence strength of WC and SC groups are weak and the liver cells array disorder and liver rope structures fuzzy. Anal 'sis from statistical data, comparing WC-group, IRS-1express in liver ce lls descend of SC-group and ascend of XYS-group. Comparing SC-group, IRS- 1express in liver cells are ascend of S+LXYS and S+HXYS groups, between the S+HXYS group is clearer.
conelusions
1Chronic-psychological-stress can make more serious hyperglycemia f or T2DM, reduce insuline sensitivity, raise TG、CHO,reduce the body weight. XYS-power'effect lies in the following:one aspect can descend the gluco se,the second can resist the hyperglycemia resulted from Chronic-psycholo gical-stress, ascend insuline sensitivity, reduce TG、CHO、LDL-C, ascend HDL-C, and reduce hyperlipidemia and body weight becaused of Chronic-psychol ogical-stress. It's effect mechanism might have increased insuline sensit ivity to correct the sugar-fat disorder.
2Because of the variegation and heterogeneity of T2DM heredity, the re is not proper, comprehensive, suitable technology and heredity analysis methods for gene-heredity diseases. The study selects89related gene of T2DM, uses PCR microchip technology to probe the influence of Chronic-psyc hological-stress to IR of T2DM and Xiao-Yao-San effect machnism. Result:c omparing to WC-group,there is7genes ascending,6genes descending of SC-group; there is8genes ascending,3genes descending of XYS-group; among FOXC2、Ifng、Slc2a4are their common ascending genes. Aqp2、Pfkfb3. Tnfrsf1b are their common descending genes.So, Chronic-psychological-stress can adjust related genes express and make more serious IR of T2DM. Xiao-Yao-San can prevent and cure diabetes and improve IR, Which it is might relate d to genes express.
3Chronic-psychological-stress can increase GR express in insulin ce11of T2DM mouses.But Xiao-Yao-San can reduce GR express in the insulin c ell of T2DM. S+LXYS and S+HXYS groups all can reduce GR express in insulin cell of Chronic-psychological-stress T2DM, and S+HDL-C group's effect is better than S+LXYS group. But GR express in liver cell of SC-group is desc end, that can relate to liver cell death resulted from long-time Chronic-p sychological-stress.GR express in liver cell of S+LXYS and S+HXYS groups are ascend, and S+HXYS group's effect is better than S+LXYS group, which it might be Xiao-Yao-San make better liver cell damage becaused of Chronic-psychol ogical-stress.
4Chronic-psychological-stress can decrease IRS-1express in insulin cell of T2DM mouses.But Xiao-Yao-San ascend IRS-1express in insulin cel l and liver cell of T2DM mouses, that the result is the same to the gene e xpress. The result is to show that inhibiting IRS-1express in insulin ce ll,reducing insulin secretion, and inhibiting IRS-1express in liver cell, reducing liver glycogen's compound might be important effect mechanism of IR of T2DM mouses resulted from Chronic-psychological-stress. Xiao-Yao-S an can ascend IRS-1express in insulin and liver cells, increase insulin s ectetion, increase liver glycogen's compound to reduce glucose and relieve insulin resistant.
引文
[1]Yang WY,Lu JM,Weng JP,et al.Prevalence of Diabetes among Men and Women in China [J].The New England Journal ofMedicine,2010,362:1090-1101.
[2]Helz JW,Templefon B.Evidence of the role of psychosocial factors indiabetes mellitus:a review[J].Am J Psychiatry,1990,147(10):1275-1282.
[3]Chandak GR, Janipalli CS, Bhaskar S, et al. Common variants in the TCF7L2 gene are strongly associated with type 2 diabetes mellitus in the Indian population [J]. Diabetologia,2007,50(1):63-67.
[4]Tong Y,Lin Y,Zhang Y,et al.Association between TCF7L2 genepolymorphisms and susceptibility to type2 diabetes mellitus:a largeHuman Genome Epidemiology(HuGE) review and meta-analysis[J].BMC Med Genet,2009,19:10-15.
[5]Sladek R,Rocheleau G,Rung J.A genome-wide association study identifies novel risk loci for type 2 diabetes[J].Nature,2007,7130:881-885.
[6]Lyssenko V,Lupi R,Marchetti P,et al.Mechanisms by which common variants in the TCF7L2 gene increase risk of type 2 diabetes[J].J Clin Invest,2007,117(8):2155-2163.
[7]Silva Xavier GD,Loder MK,McDonald A,et al.TCF7L2 regulates late events in insulin secretion from pancreatic islet Beta-cells[J]Diabetes,2009,58(4):894-905.
[8]洪靖.李光伟.IRS-1基因与肥胖、2型糖尿病.中日友好医院学报,2003，17(2)：107-109.
[9]FedericiM,HribalML,RanalliM,etal.The common Arg972polymorphism in insulin receptor substrate-1 causes apoptosisof human pancreatic islets[J].FASEB,2001,15(1):22-24.
[10]Chimienti F,Favier A,Seve M.ZnT-8,a pancreatic beta-cell-specific zinc transporter [J].Biometals,2005,18(4):313-317.
[11]王玉霞,张丹,吴志香,等.PPARγ2基因Prol2A1a多态性与超重和肥胖的相关性[J].现代预防医学2010.36(20)：56-60.
[12]Chu ZH, Zhang J, Tian SK, et al. Correlation between SNP308 of TNF-α Gene and Susceptibility to Type 2 Diabetes in Xinjiang Uygur Population [J].Progress in Modern Biomedicine, 2009,6:1098-1100.
[13]Kem PA, Saghizadeh M, Ong JM, et al.The expression of tumor necrosis factor in human adipose tissue.Regulation by obe necrosis factor in human adipose tissue.Regulation by obesity.weight loss,and relationship to lipoprote in lipase[J].Clin Invest,1995,95(5):2111-2119.
[14]黄文凡,文秀英.余杰.FOXO1转录因子在肝脏糖脂代谢中的作用[J].中西医结合研究.2010,2(6)：317-320.
[15]Farmer SR.The forkhead transcription factor FOXO1:Apossible link between obesity and insulin resistance[J].Molcell,2003,11(1):6-8.
[16]Nakae J,Kitamura T,Kitamure Y,et al.The forkhead transcription factor FOXO1 regulates adipocyte differentiation [J].Dev Cell,2003,4(1):119-129.
[17]赵萸,罗敏.脂肪细胞分化的调控及与2型糖尿病的关系[J].国外医学：内分泌学分册,2004.24(1)：3-6.
[18]Cerderberg A,Gronning LM,Ahren B,et al.FOXC2 is a winged helix gene that counteracts obesity,hypertriglyceridemia,and diet-induced insulin resistance[J].Cell.2001,106:563-573.
[19]Kovacs P,Lehn Stefana,Stumvollm,et al.Genetic variation in the human winged helix/forkhead transcription factor gene FOXC2 in Pima Indians[J].Diabetes,2003,52(5):1292-1295.
[20]Jasonk, KIM, HyoJeongkim,et al.Adipocyte specific overexpression of FOXC2 prevents dietinduced increases in intramuscular fatty acyl coa and insulin resistance[J].Diabetes,2005,54:1657-1663.
[21]Huang S,Czech MP.The GLUT4 glucose transporter[J].Cell Metab,2007,5(4):237-252.
[22]Friedel S,Antwerpen B.Hoch A,et al.Glucose transporter 4 gene:association studies pertain-ing to alleles oftwo polymorphisms in extremely obese children and adolescents and in normal and underweight controls [J].Ann NY AcadSci,2002,967(6):554-557.
[23]Bodhini D.Radha V,Ghosh S,et al.GLUT4 gene polymorphisms and their association with type 2 diabetes insouth Indians [J].Diabetes TechnolTher,2011,13(9):913-920.
[24]C.Y.Zhang,L.E.Parton,C.P.Ye, S.Krauss, R. Shen, C.T.Lin, C.T.Lin, J. A. PorcoJr, B.B, Lowell.Genipin inhibits UCP2-mediated protonleak and acutely reverses obesity-and hinghglu-coseinduced βcell dysfunction in isolated pancreatic islets [J].Cell Metabolism,2006:417-427.
[25]Derzbach L,Bokodi G,Treszl A,et al.Selectin polymorphisms andperinatal morbidity in low-birthweight infants[J].ActaPaediatr,2006,95(10):1213-1217.
[26]Kajantine E.Osmond C.Barker DJ.et al.Preterm birth a risk factor for type 2 diabetes? The Helsinki birth cohort study[J].Diabetes Care,2010,33(12):2623-2625.
[27]Stavarachi M.Panduru NM.Serafinceanu C,et al.Investigation of P213S SELL gene polymorphism in type 2 diabetes mellitus and related end stage renal disease.A case-control study[J].Rom J MorpholEmbryol.2011,52(suppl):995-998.
[28]徐三妮,刘静,郭茜,等.甘肃东乡族L-选择素P213S基因多态性与2型糖尿病的相关性研究[J].临床荟萃,2009,24(16)：1384-1387.
[29]Diamond J.The double puzzle of diabetes.Nature.2003.42(3):599-602.
[30]Zimmet P.Alberti K G.Shaw J.Global and societal implicationsof the diabetes epidemic. Nature,2001,414(6865):782-787.
[31]富振英,马林茂,王克安,等.我国2型糖尿病的流行特征[J].中国慢性病预防与控制,1999,7(3)：125-128.
[32]Wang Y,Rimm EB.Stampfer MJ.et al.Comparison of abdominal adiposity and overall obesity in predicting risk of type 2 diabetesamong men[J].Arn J ClinNutr.2005.81(3):555-560.
[33]张维忠.肥胖、2型糖尿病与高血压[J].中华内科杂志,2002,41(4)：284-288.
[34]张毓洪,陈启众,贺宝福,等.银川市社区2型糖尿病危险因素病例对照研究[J].宁夏医学杂志,2006,28(2)：95-97.
[35]MizockBA.Alterationsincarbohydratemetabolismduringstress:areviewoftheliterature [J].AmJMed,1995.98(1):75-84.
36.石立,钟历勇,舒毅.应激对2型糖尿病患者神经内分泌相关激素的影响[J].现代医学,2006,34(5)：321-323.
[37]Bjorntorp P.endocrine abnormalities of obesity. Metabolism,1995,44(9suppl 3):21-23.
[38]Bjorntorp P.The origins and consequences of obesity.Diabetes.Ciba-Found-Symb.1999,201:68-80.
[39]Bjorntorp P.stress and cardiovascular disease.Acta-Physiol-Scand-Suppl.1997,640:144-148.
[40]ReavenGM.Role of insulin resistance in human disease[J].Diabetes.1998.37(11):1595-1607.
[41]Ron D,Walter P.Signal integration in the endoplasmic reticulumunfolded protein response[J].Nat Rev Mol Cell Biol 2007.8(7):519-529.
[42]Shen N.Yu X,Pan FY.et al.An early response transcription factor,Egr-1,enhances insulin resistance in type 2 diabetes with chronic hyperinsulinism[J].J Biol Chem.2011.286:14508-14515.
[43]BjorntorpP.Holm G,Rewen-L.etal.Hypothalamic arousalinsulin resistance and type2 diabetes mellitus[J].Diabet-Med.1999.16(5):373-383.
[44]Loytsker J.Aiello JR.Internet addiction and its personality correlatesPaper presented at the annual meeting of the EasternPsychological Association Washington,DC,April 11,1997.
[45]Greenfild DN.The Nature of Internet Addiction:PsychologicalFactors in Compulsive Internet Use.Paper presented at the 1999meetings of the American Psychological Association,BostonMassachusetts,August 20,1999.
[46]Anderson KJ.Internet use among college students:An exploratory study. The Journal of American College Health,2001,50(1):21-26.
[47]Surwit RS,Schneider MS.Role of stress in the etiology and treatment of diabetes mellitus[J].Psychosom Med,1993,55:380-386.
[48]Mikat EM,Hackel DB,Cruz PT and Lebovitz HE.Lowered glucose tolerance in the sand rat (psammonysobesus)resulting from esophageal incubation[J].Proc SocExp Biol Med,1972,139:1390-1395.
[49]BJorntorp P.Endocrine abnormalities in human obesity[J].Metabolism,1995.44(9)Suppl:321-325.
[50]Surwit RS,Feinglos MN.Livingston EG,Kuhn CM,McCubbin J A.Behavioral Manipulantion of the diabetic phenotype in ob/ob mice[J].Diabetes,1984,33:616-623.
[51]Surwit RS,McCubbin J A,Kuhn CM,McGee D,Gerstenfeld DA,Feinglos MN.Alprazolam reduces stress hyperglycemia in ob/ob mice[J].Psychosom,Med.1986,48:278-284.
[52]DunbarHF.Psychiatric aspect of medicalproblems[J].AmJ.Psychiatry,1936,(93):649-656.
[53]Surwit RS,Schneider MS,Feinglos MN.Stress and diabetes mellitus[J].Diabetes Care,1992,15(10): 1413-1419.
[54]SurwitRS.Feinglos MN.Stress and autonomic nervous system in type 2 diabetes.diabetes Care [J], 1988.11(1):83-89.
[55]姚树桥,蔡伟雄，戴晓阳等.实验性应激对链脲佐菌素诱导鼠血糖增高的机理研究.中华医学杂志.1999,79：332-334.
[56]龚耀先，姚树桥,戴晓阳等.心理社会因素在糖尿病发生过程中的作用及机理研究I理论框架[J].中国临床心理学杂志，1997,5(2)：69-73
[57]姚树桥,戴晓阳,高北陵.应激在糖尿病发生中的作用及机理[J].中国临床心理学杂志.1995.3(4)：249-252.
[58]戴晓阳,周智广,姚树桥等.心理社会因素在糖尿病发生过程中的作用及机理研究Ⅳ.Ⅱ型糖尿病发生的危险因素分析[J].中国临床心理学杂志1998,.6(4)：196-202.
[59]李秀平,孙静,张俊权等.慢性应激对大鼠葡萄糖耐量及胰岛素抵抗的影响[J].中国医药导报.2007,4(28)：25-26.
[60]高北陵，戴晓阳,姚树桥等.紧张性生活事件在人类Ⅱ型糖尿病发生中的作用[J].中国临床心理学杂志.1997.5(3)：134-138.
[61]George, Chrousos.应激与应激系统紊乱的概念—概述身体的和行为的内环境稳定[J].美国医学杂志中文版，1992，38(5)：290-295.
[62]Surwit RS, Schneider MS. Role of stress in the etiology and treatment of diabetes mellitus[J]. Psychosomatic Medicine.1993,55(6):380-393.
[63]王丽,薛志敏,姚树桥等.心理应激对2型糖尿病人糖代谢影响的对照研究[J].中国临床心理医学杂志.2000，8(2)：98-101.
[64]朱熊兆,陈晓岗,姚树桥等.糖尿病患者的焦虑情绪及其对糖代谢控制的影响[J].中国行为医学科学，2002.11(3)：163-164
[65]高北陵，龚耀先,戴晓阳等.心理社会因素在糖尿病发生过程中的作用及机理研究Ⅱ紧张性生活事件对人类2型糖尿病发生影响的初步研究[J].中国临床心理学杂志.1997.5(3)：134-139.
[66]蔡太生,龚耀先,高北陵等.生活事件在2型糖尿病发病中作用的追踪研究[J].中国临床心理学杂志,2002,10(4)：249-251.
[67]Von Bochmer H, Sarukhan A.GAD. a single autoimmune for diabetes[J]. Science.1999, 284:1135-1143.
[68]Yoon JW, Yoon CS, Lim HW, etal.Control of autoimmune diabetes NOD mice by GAD expression or suppression inβ-Cell[J]. Science.1999.284:2183-2187.
[69]Chan O, Chan S, Inouye K et all Molecular regulation of the hypothalamo-p ituitary-adrenal axis in strep tozotocin induced diabetes:effects of insulin treatment[J]. J Endocrinolo2gy,2001,142: 4872-4879
[70]ChanO, Inouye K, VaranicM et al. Hyperactivation of the hypothalamo-pituitary-adrenocor--tical axis in strep tozotocin diabetes is associated with reduced stress responsivenessand decreased pituitary and adrenal sensitivity[J]. J Endocrinology,2002,143:1761-768.
[71]Amaldi G, Angeli A, Atkinson AB et al. Diagnosis and complications of Cushing's syndrome:a consensus statement[J]. J Clin Endocrinol Metab,2003,88:5593-5602.
[72]Tauchmanova LI. Rossi R. Biondi B et al. Patients with subclinical Cushing's syndrome due to adrenal adenoma have increased cardiovascular risk[J]. J Clin Endocrinol Metab,2002,87:4812-4878.
[73]BjorntorpP, Holm G, Rewen-L. etal. Hypothalamic arousalinsulin resistance and type2 diabetes mellitus[J]. Diabet-Med,1999,16(5):373-383.
[74]BJorntorp P.Neuroendocrine abonormalities in human obesity[J].Metabolism,1995,44(2) Suppl 2: 38-46.
[75]BJorntorp P. Endocrine abnormalities of obesity[J].Metabolism,1995.44(9)Suppl 2:38-43.
[76]BJorntorp P.Visceral obesity:A"Civilization syndrome"[J].Obes Res.1993.1:206-210.
[77]Kissebah AH.Peiris AN.Biology regional body fat distribution.Relationship to non-insulin-dependent diabetes mellitus[J].Diabetes Metab Rev,1989,31(5):83-87.
[78]Surwit RS.Feinglos MN.Stress and automonomic nervous system in type Ⅱ diabetes:ahypothesis.Diabetes Care[J].1988.11 (1):83-90.
[79]. Ron D, Walter P. Signal integration in the endoplasmic reticulumunfolded protein response[J]. Nat Rev Mol Cell Biol 2007,8(7):519-529.
[80]Shen N, Yu X, Pan FY, et al. An early response transcription factor. Egr-1. enhances insulin resistance in type 2 diabetes with chronic hyperinsulinism[J]. J BiolChem,2011,286:14508-14515
[81]孙万森，吴喜利，杨成志.消渴病病机循证研究[C].第八次全国中医糖尿病学术大会论文汇编,2005：27-29.
[82]李渊何.糖尿病中医病机再探)试论“五脏皆柔弱”为糖尿病基本病机[J].天津中医药.2004.21(3)：216-217.
[83]翁銮坤.脾虚在糖尿病发病机制中的地位[J].中华中医药学刊.2007.25(10)：2158-2160.
[84]郭军会,吕爱平.中医脾肾与糖尿病发病关系初探[J].辽宁中医药大学学报.2009，11(8)：81-82.
[85]吴峥嫌.糖尿病中医病机浅淡[J].新中医，2008, 40(3)：100-101.
[86]邹时祯.2型糖尿病的中医核心病机探析[J].亚太传统医药，2009.5(5)：5-6.
[87]郑红.试论脾失健运是消渴病发病的内在依据[J].湖北中医医杂志.2009.30(2)：30-31
[88]王明选.旷云祥.“从肝论治”学说治疗糖尿病举偶[J].贵阳中医学院学报.2011.33(5)：117-119
[89]韩乐兵，陶毅.糖尿病从肝胆论治初探[J].辽宁中医杂志.2003.30(8)：611-612.
[90]刘绪银.石海澄.王笃高等.论肝与消渴[J].湖南中医药导报.2000.6(3)：13-14.
[91]李道本，工智明.从肝论治防治胰岛素抵抗治疗2型糖尿病的理论探讨[J].中国中西医结合消化杂 志,2001,9(3)：153-156.
[92]王明选,旷云祥.“从肝论治”学说治疗糖尿病举偶[J].贵阳中医学院学报,2011,33(5)：117-119
[93]马鑫,李杰.糖尿病的心身并治[J].天津医学，,2000,17(6)：39-42.
[94]高绍芳,赵克景,杜立建.浅谈糖尿病从肝论治[J].现代中西医结合杂志,2006,15(18)：2151-2153.
[95]陈炳.中医从肝论治糖尿病的研究[J].中国民康医学,2006,18(12)：994-996.
[96]马万千.论情志失调与消渴病[J].中国误诊学杂志,2003，3(5)：772-773.
[97]王钢柱.中医疏肝调气法治疗糖尿病.北京：人民卫生出版社，2001.4.
[98]邹如政.糖尿病从肝论治[J].中国医药学报,1998,35(1)：17-23.
[99]左嫒媛,杨云.从肝失疏泄谈糖尿病的发生[J].云南中医学院学报,2003，26(1)：22-23.
[100]金殿青.试论糖尿病从肝论治[J].中医药研究，1991,(2)：64-67.
[101]安俊义.任佑才应用疏肝法治疗消渴病经验[J].山西中医,1995,15(6)：7-8.
[102]段尚勤.消渴之治勿忘调肝[J].光明中医，1996,(2)：15-16.
[103]孙传庆.抑肝法为主治疗糖尿病[J].中医药研究,1990,(1)：30-33.
[104]李小杵.试论糖尿病从肝论治[J].中医药研究，1996，(3)：24-27.
[105]秦宗昌.试论糖尿病从肝辨治[J].新中医.1993，(4)：13-14.
[106]于志强.酸泻肝木法治疗消渴[J].四川中医.1994,(2)：24-25.
[107]张挺群.2080例糖尿病患者症候与血糖关系分析[J].中医杂志.1996,37(10)：617-619.
[108]张世筠，杜宝俊，周群清等.中医肝证与糖尿病患病关系的研究[J].中国中医药信息杂志.2003.(10)7：90-91.
[109]苏浩.仝小林,王皖洁.仝小林教授治疗糖尿病学术观点和经验[J].中国医药指南,2008,6(24)：198-199
[110]杨辰华.刘完素玄府气液理论与2型糖尿病病机及治疗的相关性探讨[J].中医研究,2009,22(1)：3-6
[111]魏桂梅.从痰瘀论治胰岛素抵抗2型糖尿病70例[J].甘肃中医,2009，22(1)：34-35
[112]江南，刘铜华.胰岛素抵抗的中医病机探讨[J].中国中医基础医学杂志.2006.12(9)：691-692
[113]纪云西,蒋历.胰岛素抵抗的中医病机实质探析[J].辽宁中医药大学学报.2008,10(3)：23-24
[114]季雁浩.从肝论治糖尿病[J].中医药研究，1993.34(1)：21-23.
[115]江武.消渴从肝论治刍议[J].中国中医药信息杂志.1996.28(11)：9-12.
[116]王智明,魏子孝.从肝论治消渴(糖尿病)的临床体会[J].中国中医基础医学杂志,2001.7(3)：65-66
[117]杨家景.糖尿病从肝论治[J].福建中医药.1997.35(6)：28-31.
[118]郭善军.逍遥降糖饮治疗糖尿病60例[J].北京中医学院学报.1991.30(3)：36-38.
[119]孟翠霞.滋水清肝饮加减治疗糖尿病38例[J].湖南中医学院学报.1995，(2)：18-20.
[120]杜丹,张文芳.刘天胤等.龙胆泻肝汤在治疗肥胖型消渴中的应用[J].中医药学报.1995.(4)：45-47.
[121]杨晓晖.四逆散加味治疗消渴病肝气郁结证[J].上海中医药杂志.1997.35(5)：11-13.
[122]刘素荣.刘桂芝.孙丰雷等.糖尿病从肝论治的理论探讨[J].山东中医药大学学报.1999(5)：340-341.
[123]倪海祥.刘刚，罗苏生等.从肝论治2型糖尿病的临床研究[J].中国中西医结合杂志，2000.20(8)：577-579
[124]鞠祥宗.韦淑萍.从肝论治糖尿病40例[J].陕西中医.2003.24(3)：201-202.
[125]周潮,李晓哲，单友琴等.降糖煎治疗II型糖尿病的临床与实验研究[J].光明中医杂志,1996,28(6)：51-54
[126]周劲刚.2型糖尿病从肝论治[J].四川中医.2012，30(4)：35-36
[127]徐志伟,敖海清,王文竹等.逍遥散对慢性心理应激损伤大鼠海马神经元的影响.中药理与临床,，2005,(21)1：5-8.
[128]徐志伟,敖海清,王文竹等.逍遥散对应激大鼠海马CA3区突触体结构可塑性的影响.中国行为医学科学杂志,2006,(15)2：102-106
[129]敖海清，徐志伟,苏俊芳等.柴胡疏肝散及逍遥散对慢性心理应激大鼠血清皮质酮及胃肠激素的影响.中药新药及临床药理,2007,(18)4：288-291.
[130]敖海清,徐志伟,富文俊等.逍遥散对慢性应激大鼠下丘脑-垂体-肾上腺轴调控作用机制的初步研究.中华行为医学与脑科学杂志.2010, (19)8：680-682.
[131]李光伟，潘孝仁,]Lillioja S等.检测人群胰岛素敏感性的一项新指数.中华内科杂志，1993，32(10)：656-660.
[132]Zimmet P,Alberti KG,Shaw J.Global and societalimplications of the diabetes epidemic [J].Nature,2001.414(6865):782-780.
[133]De Fronzo RA.Tobin J D, Andres R. Glucose clamp technique:a method for quantifying insulin secretion and resistance [J]. Am JJOURNAL OF PRACTICAL DIABETOLOGY Vol.7 NoPhys-iol 1979.237 (3):214-223.
[134]Matthews D R, Hosker J P.Rudenski A S.et al.Homeostasismodel assessment:insulin resist-ance and beta-cell function from fastingplasma glucose and insulin concentrations in man.Dia-betologia [J].1985.28(7):412-419.
[135]Ravipat IG,Aronow WS,Ahn C. et al.Association of hemoglobin Alc levelwith the severity of coronary artery disease in patients with diabetes mellitus[J].Am J Cardiol.2006,97(7):968-969.
[136]王喜,郑华丽.120例糖尿病患者血脂血糖的监测与分析[J].中国医药科学.2011，1(12)：111-112.
[137]McCann BS.Benjamin GA,W ilkinson CW. et al. Plasma lipid concentrationsduring episodic occupa- tional stress[J].Ann BehavMed,1999,21(2):103-110.
[138]Gottdiener JS,KopWJ.HausnerE, eta.l Effects ofmentalstress on flow-mediated brachial arterial dilation and influence ofbehavioral factors and hypercho-lesterolemia in subjects without cardio-vasculardisease[J].Am JCardiol 2003.92(6):687-691.
[139]Bacon SL,Ring C.Lip GY,et al.Increases in lipids and immune cells in response to exercise andmental stress in patientswith suspected coronary arterydisease:effects of adjustment for shifts in plasma volume[J].Biol Psychol.2004.65(3):237-250.
[140]Bachen EA,MuldoonMF,MatthewsKA.eta.l Effects ofhemoconcentration andsympathetic activation on serum lipid responses to briefmental stress[J]. PsychosomMed.2002.64(4):587-594.
[141]Chandak GR, Janipalli CS, Bhaskar Set al. Common variants in the TCF7L2 gene are strongly associated with type 2 diabetes mellitus in the Indian population [J]. Diabetologia,2007,50(1):63-67.
[142]王旭东.丁选平.2型糖尿病相关基因研究进展.中国优生与遗传杂志，2006，14(1)：4-5.
[143]罗敏.2型糖尿病相关基因的研究现状及展望.国外医学内分泌学分册，2002.22(3)：141-145.
[144]Mangion J. Rahman N. Mansour S. et al. A genefor lymphedema- distichiasis maps to 16q24.3 [J]. Am JHum Genet,1999,65:427-432.
[145]Cerderberg A, Gronning LM, Ahren Bet al. FOXC2 is a winged helix gene that counteracts obesity, hypertriglyceridemia, and diet-induced insulin resistance[J].Cell.2001.106:563-573.
[146]Jasonk Kim,Hyo-Jeong Kim et al.Adipocyte-specific overexpression of FOXC2 prevents diet-induced increases in intramuscular fatty acyl coa and insulin resistance[J].Diabetes,2005.54:1657-1663.
[147]YangX, SmithU.Decreased adipose tissue FOXC2 expression in insulin resistance-impaired precursor cell commitment to bothwhite andbrown adipose differentiation [C]. ADA 63rd Scientific Sessions,2003:381.
[148]DahleMK,Gronning LM, Cederberg A, et a.l Mechanisms of FOXC2and FOXD1-mediated regulation of the RI alpha subunit of campdependentprotein kinase include release of transcript-tional repressionand activation by protein kinaseB alpha and cAMP[J]. JBiolChem.2002,277(25): 22902-22908.
[149]Di Gregorio GB, Westergren R. Enerback S. et a.l Expression ofFOXC2 in adipose and muscle and its association with whole bodyinsulin sensitivity[J].Am J Physiol EndocrinolMetab.2004,287. (4):E709-E803.
[150]Kim JK.Kim HJ, Park S, et a.l Adipocyte-specific overexpression ofFOXC2 prevents diet-induce increases in intramuscular fatty acylCoA and insulin resistance[J].Diabetes,2005,54:1657-1673.
[151]YuC,ChenGW. ZhangD, eta.lMechanism bywhich fatty acids inhibit insulin activation of insulin receptor substrate-1(IRS-1)-associated phosphatidylinositol 3-kinase activity in muscle[J], J BiolChem, 2002.277:50230-50236.
[152]De Souza CT. Araujo EP, Stoppiglia LF. et al. Inhibition ofUCP2 expression reverses dietinduced diabetes mellitus byeffects on both insulin secretion and action[J]. FASEB J.2007,21(4):1153-1163.
[153]Pi J. Bai Y. Daniel KW. et al. Persistent oxidative stress dueto absence of uncoupling protein 2 associated with impairedpancreatic beta-cell function[J].Endocrinology.2009,150 (7):3040-3048.
[154]Affourtit C, Jastroch M. Brand MD. Uncoupling protein-2attenuates glucose-stimulated insulin secretion in INS-1Einsulinoma cells by lowering mitochondrial reactive oxygenspecies[J].Free Radic Biol Med,2011.50(5):609-616.
[155]Divakaruni AS, Brand MD.The regulation and physiology ofmitochondrial proton leak [J]. Physiology (Bethesda).2011.26(3):192-205.
[156]Leloup C, Tourrel-Cuzin C, Magnan C. et al. Mitochondrialreactive oxygen species are obligatory signals for glucose-induced insulin secretion[J]. Diabetes.2009.58 (3):673-681.
[157]Lablanche S.Cottet-Rousselle C.Lamarche Fet al.Protectionof pancreatic INS-1 beta-cells from glucose and fructose-induced cell death by inhibiting mitochondrial permeabilitytransition with cyclosporin A or metformin[J]. Cell DeathDis.2011.2(1):134-145.
[158]Pi J. Collins S. Reactive oxygen species and uncouplingprotein 2 in pancreatic beta-cell functional]. Diabetes ObesMetab.2010.12(Suppl 2):141-148.
[159]Harmon JS. Stein R. Robertson RP. Oxidative stress-mediated, post-translational loss of MafA protein as acontributing mechanism to loss of insulin gene expression inglucotoxic beta cells[J]. J Biol Chem,2005,280(12):11107-11113.
[160]Robertson RP. Chronic oxidative stress as a centralmechanism for glucose toxicity in pancreatic islet beta cells indiabetes[J]. J Biol Chem,2004,279(41):42351-42354.
[161]Pi J, Bai Y, Daniel KW, et al. Persistent oxidative stress dueto absence of uncoupling protein 2 associated with impairedpancreatic beta-cell function [J]. Endocrinology,2009,150(7):3040-3048.
[162]Affourtit C, Jastroch M, Brand MD. Uncoupling protein-2attenuates glucose-stimulated insulin secretion in INS-lEinsulinoma cells by lowering mitochondrial reactive oxygenspecies [J]. Free Radic Biol Med,2011,50(5):609-616.
[163]BraissantO,FouflleF,Scotto C et al. Differential expression of peroxisome proliferator--activated receptors (PPARs):tissue distribution of PPAR-alpha,-beta, and-gamma in the adult rat [J]. Endocrinology,1996,137(1):354-366.
[164]Akiyama T E, Nicol CJ, Fievet C et al. Peroxisome proliferator-activated receptor-alpha regulates lipid homeostasis, but is not associated with obesity studies with congenic mouse lines [J]. J Bi1 Chem, 2001,276(42):39088-39093.
[165]Li P,Zhu Z,Lu Y,et al. Metabolic and cellular plasticity in white adipose tissue O:role of peroxisome proliferator-activated receptor [J].Am J Physiol Endocrinol Metab,2005.289(4):E617-E626.
[166]Anandharajan R, Pathmanathan K, Shankernarayanan N P, et al. Upregulation of Glut-4 and PPARgamma by an isoflavone from Pterocarpus marsupium on L6 myotubes:a possible mechanism of action [J]. Ethnopharmacol,2005,97(2):253-260.
[167]Hegele RA.Monogenic forms of insulin resistance:apertures that expose the common metabolicsyndrome [J]. Trends Endocrinol Metab,2003,14(8):371-377.
[168]Enomoto N.Takei Y. Hirose M, et al. Prevention of ethanol induced liver injury in rats by an a gonist of peroxisome proliferators-activated receptor-gamma pioglitazone [J].Pharmacol ExpT--her.2003, 306(3):846-854.
[169]Barens PJ, Adock IM. Anti-inflammatory actions of steroids; molecularmechansims[J].Trends Pharmacol Sci,1993.14:436-441.
[170]Delaunay F. KhanA, Cintr A, etal. Pancreatic beta cells are important targets for the diabetogenic effects of glucocorticoids [J]. J Clin Invest,1997,100:2094-2098.
[171]Ling ZC,KhanA.DelaunyF,et al. Increased glucocorticoid sensitivity in islet beta-cells:effectson glucose-6-phosephatase,glucose cycling and insulin release [J]. Diabetologia,1998.41:634-639.
[172]Itoi K, Jiang YQ, Iwasaki Y. et al. Regulatory mechanism ofcorticotrophin-releasing hormone and va--sopressin gene expression inthe hypothalamus[J]. Neuroendocrinol,2004,16(4):348-355.
[173]ChanO.Inouye K.AkiravEM,et al.Hyperglycemia does notincrease basal hypothalamo-pituitary-adrenal activity in diabetes butit does impair the HPAresponse to insulin-induced hypoglycemia[J].Endocrinology.2005.146(3):1382-1390.
[174]崔俊芳，张如意.2型糖尿病患者的血浆皮质醇水平的变化及与代谢指标的相关分析[J].宁夏医学杂志,2010.32(6)：535-537.
[175]Evans SB.Wilkinson CW.Gronbeck P,et al.Inactivation of the DMHselectively inhibits the ACTH and corticosterone responses tohypoglycemia[J].Am J Physiol Regul Integr Comp Physiol,2004, 286(1):123-128.
[176]NyirendaMJ, LindsayRS.KenyonCJ.etal. Glucocorticoidexposure inlate gestation permanently programsrathepatic phosphoenolpyruvatecarboxy kinaseandglucocorticoidreceptorexpressionandcauses glucosein tolerance adultoffspring [J].JClinlnvest,1998.101:2174-2181.
[177]李芳萍,张四清,王斐等.2型糖尿病和非酒精性脂肪肝病的胰岛素抵抗研究[J].新医学,2009.40(7)：427-30.
[178]刘淑娥,肖丹，肝源性糖尿病临床分析[J].中日友好医院学报,2003,17(3)：153-154.
[179]Myers G M Jr, Sun X J,et al. The IRS-1 signaling system. TIBS,1994,19(7):289-293
[180]唐伟,朱剑,武晓泓等.胰岛素信号转导及其调节机制[J].实用糖尿病杂志,2005,1(3)：43-49.
[181]周云枫,吴勇,欧阳静萍.黄芪多糖对2型糖尿病大鼠肾组织胰岛素信号转导的影响[J].武汉大学学报,2005,26(2)：139-143.
[182]Aspinwall CA, Qian WJ, Roper MG, et al. Roles of insulin receptorSubstrate-1. phosphatidylinosi--tol 3-kinase, and release of intracellular Ca2+ stores in insulin-stimulated insulin secretion inβ-cells[J]. J BiolChem.2000,275(29):22331-22338.
[183]张妍，袁莉.p细胞胰岛素受体信号系统与p细胞功能[J].国外医学内分泌学分册，2005,25(4)：259-261.
[184]裴海成.高浓度游离脂肪酸对胰岛素受体后信号转导的下降调节[J].第二军医大学学报，2003,24(5)：569-573.
[185]Yuan L.Ziegter R.Hamann A.Meffomdn modulates insulin post-receptorsignalingtrans- duction in chronically insulin-treated HepG2 cells[J]. Acta Pharmacol Sin.2003,24(1):55-60.
[186叶‘华.2型糖尿病患者及其一级亲属和非糖尿病者骨骼肌的胰岛索信号传导[J].中国糖尿病杂志，2003，11(2)：151-152.
[187]彭定琼，高妍,陈宇等.OLETF大鼠肝脏、肌肉、脂肪组织中胰岛素受体底物1的蛋白表达[J].中华内分泌代谢杂志.001，17(5)：288-289.
[188]张中成，刘志诚.2型糖尿病胰岛素受体后水平研究现状[J].医学综述，2002，8(3)：166-167.
[2]Helz JW,Templefon B.Evidence of the role of psychosocial factors indiabetes mellitus:a review[J].Am J Psychiatry,1990,147(10):1275-1282.
[3]Chandak GR, Janipalli CS, Bhaskar S, et al. Common variants in the TCF7L2 gene are strongly associated with type 2 diabetes mellitus in the Indian population [J]. Diabetologia,2007,50(1):63-67.
[4]Tong Y,Lin Y,Zhang Y,et al.Association between TCF7L2 genepolymorphisms and susceptibility to type2 diabetes mellitus:a largeHuman Genome Epidemiology(HuGE) review and meta-analysis[J].BMC Med Genet,2009,19:10-15.
[5]Sladek R,Rocheleau G,Rung J.A genome-wide association study identifies novel risk loci for type 2 diabetes[J].Nature,2007,7130:881-885.
[6]Lyssenko V,Lupi R,Marchetti P,et al.Mechanisms by which common variants in the TCF7L2 gene increase risk of type 2 diabetes[J].J Clin Invest,2007,117(8):2155-2163.
[7]Silva Xavier GD,Loder MK,McDonald A,et al.TCF7L2 regulates late events in insulin secretion from pancreatic islet Beta-cells[J]Diabetes,2009,58(4):894-905.
[8]洪靖.李光伟.IRS-1基因与肥胖、2型糖尿病.中日友好医院学报,2003，17(2)：107-109.
[9]FedericiM,HribalML,RanalliM,etal.The common Arg972polymorphism in insulin receptor substrate-1 causes apoptosisof human pancreatic islets[J].FASEB,2001,15(1):22-24.
[10]Chimienti F,Favier A,Seve M.ZnT-8,a pancreatic beta-cell-specific zinc transporter [J].Biometals,2005,18(4):313-317.
[11]王玉霞,张丹,吴志香,等.PPARγ2基因Prol2A1a多态性与超重和肥胖的相关性[J].现代预防医学2010.36(20)：56-60.
[12]Chu ZH, Zhang J, Tian SK, et al. Correlation between SNP308 of TNF-α Gene and Susceptibility to Type 2 Diabetes in Xinjiang Uygur Population [J].Progress in Modern Biomedicine, 2009,6:1098-1100.
[13]Kem PA, Saghizadeh M, Ong JM, et al.The expression of tumor necrosis factor in human adipose tissue.Regulation by obe necrosis factor in human adipose tissue.Regulation by obesity.weight loss,and relationship to lipoprote in lipase[J].Clin Invest,1995,95(5):2111-2119.
[14]黄文凡,文秀英.余杰.FOXO1转录因子在肝脏糖脂代谢中的作用[J].中西医结合研究.2010,2(6)：317-320.
[15]Farmer SR.The forkhead transcription factor FOXO1:Apossible link between obesity and insulin resistance[J].Molcell,2003,11(1):6-8.
[16]Nakae J,Kitamura T,Kitamure Y,et al.The forkhead transcription factor FOXO1 regulates adipocyte differentiation [J].Dev Cell,2003,4(1):119-129.
[17]赵萸,罗敏.脂肪细胞分化的调控及与2型糖尿病的关系[J].国外医学：内分泌学分册,2004.24(1)：3-6.
[18]Cerderberg A,Gronning LM,Ahren B,et al.FOXC2 is a winged helix gene that counteracts obesity,hypertriglyceridemia,and diet-induced insulin resistance[J].Cell.2001,106:563-573.
[19]Kovacs P,Lehn Stefana,Stumvollm,et al.Genetic variation in the human winged helix/forkhead transcription factor gene FOXC2 in Pima Indians[J].Diabetes,2003,52(5):1292-1295.
[20]Jasonk, KIM, HyoJeongkim,et al.Adipocyte specific overexpression of FOXC2 prevents dietinduced increases in intramuscular fatty acyl coa and insulin resistance[J].Diabetes,2005,54:1657-1663.
[21]Huang S,Czech MP.The GLUT4 glucose transporter[J].Cell Metab,2007,5(4):237-252.
[22]Friedel S,Antwerpen B.Hoch A,et al.Glucose transporter 4 gene:association studies pertain-ing to alleles oftwo polymorphisms in extremely obese children and adolescents and in normal and underweight controls [J].Ann NY AcadSci,2002,967(6):554-557.
[23]Bodhini D.Radha V,Ghosh S,et al.GLUT4 gene polymorphisms and their association with type 2 diabetes insouth Indians [J].Diabetes TechnolTher,2011,13(9):913-920.
[24]C.Y.Zhang,L.E.Parton,C.P.Ye, S.Krauss, R. Shen, C.T.Lin, C.T.Lin, J. A. PorcoJr, B.B, Lowell.Genipin inhibits UCP2-mediated protonleak and acutely reverses obesity-and hinghglu-coseinduced βcell dysfunction in isolated pancreatic islets [J].Cell Metabolism,2006:417-427.
[25]Derzbach L,Bokodi G,Treszl A,et al.Selectin polymorphisms andperinatal morbidity in low-birthweight infants[J].ActaPaediatr,2006,95(10):1213-1217.
[26]Kajantine E.Osmond C.Barker DJ.et al.Preterm birth a risk factor for type 2 diabetes? The Helsinki birth cohort study[J].Diabetes Care,2010,33(12):2623-2625.
[27]Stavarachi M.Panduru NM.Serafinceanu C,et al.Investigation of P213S SELL gene polymorphism in type 2 diabetes mellitus and related end stage renal disease.A case-control study[J].Rom J MorpholEmbryol.2011,52(suppl):995-998.
[28]徐三妮,刘静,郭茜,等.甘肃东乡族L-选择素P213S基因多态性与2型糖尿病的相关性研究[J].临床荟萃,2009,24(16)：1384-1387.
[29]Diamond J.The double puzzle of diabetes.Nature.2003.42(3):599-602.
[30]Zimmet P.Alberti K G.Shaw J.Global and societal implicationsof the diabetes epidemic. Nature,2001,414(6865):782-787.
[31]富振英,马林茂,王克安,等.我国2型糖尿病的流行特征[J].中国慢性病预防与控制,1999,7(3)：125-128.
[32]Wang Y,Rimm EB.Stampfer MJ.et al.Comparison of abdominal adiposity and overall obesity in predicting risk of type 2 diabetesamong men[J].Arn J ClinNutr.2005.81(3):555-560.
[33]张维忠.肥胖、2型糖尿病与高血压[J].中华内科杂志,2002,41(4)：284-288.
[34]张毓洪,陈启众,贺宝福,等.银川市社区2型糖尿病危险因素病例对照研究[J].宁夏医学杂志,2006,28(2)：95-97.
[35]MizockBA.Alterationsincarbohydratemetabolismduringstress:areviewoftheliterature [J].AmJMed,1995.98(1):75-84.
36.石立,钟历勇,舒毅.应激对2型糖尿病患者神经内分泌相关激素的影响[J].现代医学,2006,34(5)：321-323.
[37]Bjorntorp P.endocrine abnormalities of obesity. Metabolism,1995,44(9suppl 3):21-23.
[38]Bjorntorp P.The origins and consequences of obesity.Diabetes.Ciba-Found-Symb.1999,201:68-80.
[39]Bjorntorp P.stress and cardiovascular disease.Acta-Physiol-Scand-Suppl.1997,640:144-148.
[40]ReavenGM.Role of insulin resistance in human disease[J].Diabetes.1998.37(11):1595-1607.
[41]Ron D,Walter P.Signal integration in the endoplasmic reticulumunfolded protein response[J].Nat Rev Mol Cell Biol 2007.8(7):519-529.
[42]Shen N.Yu X,Pan FY.et al.An early response transcription factor,Egr-1,enhances insulin resistance in type 2 diabetes with chronic hyperinsulinism[J].J Biol Chem.2011.286:14508-14515.
[43]BjorntorpP.Holm G,Rewen-L.etal.Hypothalamic arousalinsulin resistance and type2 diabetes mellitus[J].Diabet-Med.1999.16(5):373-383.
[44]Loytsker J.Aiello JR.Internet addiction and its personality correlatesPaper presented at the annual meeting of the EasternPsychological Association Washington,DC,April 11,1997.
[45]Greenfild DN.The Nature of Internet Addiction:PsychologicalFactors in Compulsive Internet Use.Paper presented at the 1999meetings of the American Psychological Association,BostonMassachusetts,August 20,1999.
[46]Anderson KJ.Internet use among college students:An exploratory study. The Journal of American College Health,2001,50(1):21-26.
[47]Surwit RS,Schneider MS.Role of stress in the etiology and treatment of diabetes mellitus[J].Psychosom Med,1993,55:380-386.
[48]Mikat EM,Hackel DB,Cruz PT and Lebovitz HE.Lowered glucose tolerance in the sand rat (psammonysobesus)resulting from esophageal incubation[J].Proc SocExp Biol Med,1972,139:1390-1395.
[49]BJorntorp P.Endocrine abnormalities in human obesity[J].Metabolism,1995.44(9)Suppl:321-325.
[50]Surwit RS,Feinglos MN.Livingston EG,Kuhn CM,McCubbin J A.Behavioral Manipulantion of the diabetic phenotype in ob/ob mice[J].Diabetes,1984,33:616-623.
[51]Surwit RS,McCubbin J A,Kuhn CM,McGee D,Gerstenfeld DA,Feinglos MN.Alprazolam reduces stress hyperglycemia in ob/ob mice[J].Psychosom,Med.1986,48:278-284.
[52]DunbarHF.Psychiatric aspect of medicalproblems[J].AmJ.Psychiatry,1936,(93):649-656.
[53]Surwit RS,Schneider MS,Feinglos MN.Stress and diabetes mellitus[J].Diabetes Care,1992,15(10): 1413-1419.
[54]SurwitRS.Feinglos MN.Stress and autonomic nervous system in type 2 diabetes.diabetes Care [J], 1988.11(1):83-89.
[55]姚树桥,蔡伟雄，戴晓阳等.实验性应激对链脲佐菌素诱导鼠血糖增高的机理研究.中华医学杂志.1999,79：332-334.
[56]龚耀先，姚树桥,戴晓阳等.心理社会因素在糖尿病发生过程中的作用及机理研究I理论框架[J].中国临床心理学杂志，1997,5(2)：69-73
[57]姚树桥,戴晓阳,高北陵.应激在糖尿病发生中的作用及机理[J].中国临床心理学杂志.1995.3(4)：249-252.
[58]戴晓阳,周智广,姚树桥等.心理社会因素在糖尿病发生过程中的作用及机理研究Ⅳ.Ⅱ型糖尿病发生的危险因素分析[J].中国临床心理学杂志1998,.6(4)：196-202.
[59]李秀平,孙静,张俊权等.慢性应激对大鼠葡萄糖耐量及胰岛素抵抗的影响[J].中国医药导报.2007,4(28)：25-26.
[60]高北陵，戴晓阳,姚树桥等.紧张性生活事件在人类Ⅱ型糖尿病发生中的作用[J].中国临床心理学杂志.1997.5(3)：134-138.
[61]George, Chrousos.应激与应激系统紊乱的概念—概述身体的和行为的内环境稳定[J].美国医学杂志中文版，1992，38(5)：290-295.
[62]Surwit RS, Schneider MS. Role of stress in the etiology and treatment of diabetes mellitus[J]. Psychosomatic Medicine.1993,55(6):380-393.
[63]王丽,薛志敏,姚树桥等.心理应激对2型糖尿病人糖代谢影响的对照研究[J].中国临床心理医学杂志.2000，8(2)：98-101.
[64]朱熊兆,陈晓岗,姚树桥等.糖尿病患者的焦虑情绪及其对糖代谢控制的影响[J].中国行为医学科学，2002.11(3)：163-164
[65]高北陵，龚耀先,戴晓阳等.心理社会因素在糖尿病发生过程中的作用及机理研究Ⅱ紧张性生活事件对人类2型糖尿病发生影响的初步研究[J].中国临床心理学杂志.1997.5(3)：134-139.
[66]蔡太生,龚耀先,高北陵等.生活事件在2型糖尿病发病中作用的追踪研究[J].中国临床心理学杂志,2002,10(4)：249-251.
[67]Von Bochmer H, Sarukhan A.GAD. a single autoimmune for diabetes[J]. Science.1999, 284:1135-1143.
[68]Yoon JW, Yoon CS, Lim HW, etal.Control of autoimmune diabetes NOD mice by GAD expression or suppression inβ-Cell[J]. Science.1999.284:2183-2187.
[69]Chan O, Chan S, Inouye K et all Molecular regulation of the hypothalamo-p ituitary-adrenal axis in strep tozotocin induced diabetes:effects of insulin treatment[J]. J Endocrinolo2gy,2001,142: 4872-4879
[70]ChanO, Inouye K, VaranicM et al. Hyperactivation of the hypothalamo-pituitary-adrenocor--tical axis in strep tozotocin diabetes is associated with reduced stress responsivenessand decreased pituitary and adrenal sensitivity[J]. J Endocrinology,2002,143:1761-768.
[71]Amaldi G, Angeli A, Atkinson AB et al. Diagnosis and complications of Cushing's syndrome:a consensus statement[J]. J Clin Endocrinol Metab,2003,88:5593-5602.
[72]Tauchmanova LI. Rossi R. Biondi B et al. Patients with subclinical Cushing's syndrome due to adrenal adenoma have increased cardiovascular risk[J]. J Clin Endocrinol Metab,2002,87:4812-4878.
[73]BjorntorpP, Holm G, Rewen-L. etal. Hypothalamic arousalinsulin resistance and type2 diabetes mellitus[J]. Diabet-Med,1999,16(5):373-383.
[74]BJorntorp P.Neuroendocrine abonormalities in human obesity[J].Metabolism,1995,44(2) Suppl 2: 38-46.
[75]BJorntorp P. Endocrine abnormalities of obesity[J].Metabolism,1995.44(9)Suppl 2:38-43.
[76]BJorntorp P.Visceral obesity:A"Civilization syndrome"[J].Obes Res.1993.1:206-210.
[77]Kissebah AH.Peiris AN.Biology regional body fat distribution.Relationship to non-insulin-dependent diabetes mellitus[J].Diabetes Metab Rev,1989,31(5):83-87.
[78]Surwit RS.Feinglos MN.Stress and automonomic nervous system in type Ⅱ diabetes:ahypothesis.Diabetes Care[J].1988.11 (1):83-90.
[79]. Ron D, Walter P. Signal integration in the endoplasmic reticulumunfolded protein response[J]. Nat Rev Mol Cell Biol 2007,8(7):519-529.
[80]Shen N, Yu X, Pan FY, et al. An early response transcription factor. Egr-1. enhances insulin resistance in type 2 diabetes with chronic hyperinsulinism[J]. J BiolChem,2011,286:14508-14515
[81]孙万森，吴喜利，杨成志.消渴病病机循证研究[C].第八次全国中医糖尿病学术大会论文汇编,2005：27-29.
[82]李渊何.糖尿病中医病机再探)试论“五脏皆柔弱”为糖尿病基本病机[J].天津中医药.2004.21(3)：216-217.
[83]翁銮坤.脾虚在糖尿病发病机制中的地位[J].中华中医药学刊.2007.25(10)：2158-2160.
[84]郭军会,吕爱平.中医脾肾与糖尿病发病关系初探[J].辽宁中医药大学学报.2009，11(8)：81-82.
[85]吴峥嫌.糖尿病中医病机浅淡[J].新中医，2008, 40(3)：100-101.
[86]邹时祯.2型糖尿病的中医核心病机探析[J].亚太传统医药，2009.5(5)：5-6.
[87]郑红.试论脾失健运是消渴病发病的内在依据[J].湖北中医医杂志.2009.30(2)：30-31
[88]王明选.旷云祥.“从肝论治”学说治疗糖尿病举偶[J].贵阳中医学院学报.2011.33(5)：117-119
[89]韩乐兵，陶毅.糖尿病从肝胆论治初探[J].辽宁中医杂志.2003.30(8)：611-612.
[90]刘绪银.石海澄.王笃高等.论肝与消渴[J].湖南中医药导报.2000.6(3)：13-14.
[91]李道本，工智明.从肝论治防治胰岛素抵抗治疗2型糖尿病的理论探讨[J].中国中西医结合消化杂 志,2001,9(3)：153-156.
[92]王明选,旷云祥.“从肝论治”学说治疗糖尿病举偶[J].贵阳中医学院学报,2011,33(5)：117-119
[93]马鑫,李杰.糖尿病的心身并治[J].天津医学，,2000,17(6)：39-42.
[94]高绍芳,赵克景,杜立建.浅谈糖尿病从肝论治[J].现代中西医结合杂志,2006,15(18)：2151-2153.
[95]陈炳.中医从肝论治糖尿病的研究[J].中国民康医学,2006,18(12)：994-996.
[96]马万千.论情志失调与消渴病[J].中国误诊学杂志,2003，3(5)：772-773.
[97]王钢柱.中医疏肝调气法治疗糖尿病.北京：人民卫生出版社，2001.4.
[98]邹如政.糖尿病从肝论治[J].中国医药学报,1998,35(1)：17-23.
[99]左嫒媛,杨云.从肝失疏泄谈糖尿病的发生[J].云南中医学院学报,2003，26(1)：22-23.
[100]金殿青.试论糖尿病从肝论治[J].中医药研究，1991,(2)：64-67.
[101]安俊义.任佑才应用疏肝法治疗消渴病经验[J].山西中医,1995,15(6)：7-8.
[102]段尚勤.消渴之治勿忘调肝[J].光明中医，1996,(2)：15-16.
[103]孙传庆.抑肝法为主治疗糖尿病[J].中医药研究,1990,(1)：30-33.
[104]李小杵.试论糖尿病从肝论治[J].中医药研究，1996，(3)：24-27.
[105]秦宗昌.试论糖尿病从肝辨治[J].新中医.1993，(4)：13-14.
[106]于志强.酸泻肝木法治疗消渴[J].四川中医.1994,(2)：24-25.
[107]张挺群.2080例糖尿病患者症候与血糖关系分析[J].中医杂志.1996,37(10)：617-619.
[108]张世筠，杜宝俊，周群清等.中医肝证与糖尿病患病关系的研究[J].中国中医药信息杂志.2003.(10)7：90-91.
[109]苏浩.仝小林,王皖洁.仝小林教授治疗糖尿病学术观点和经验[J].中国医药指南,2008,6(24)：198-199
[110]杨辰华.刘完素玄府气液理论与2型糖尿病病机及治疗的相关性探讨[J].中医研究,2009,22(1)：3-6
[111]魏桂梅.从痰瘀论治胰岛素抵抗2型糖尿病70例[J].甘肃中医,2009，22(1)：34-35
[112]江南，刘铜华.胰岛素抵抗的中医病机探讨[J].中国中医基础医学杂志.2006.12(9)：691-692
[113]纪云西,蒋历.胰岛素抵抗的中医病机实质探析[J].辽宁中医药大学学报.2008,10(3)：23-24
[114]季雁浩.从肝论治糖尿病[J].中医药研究，1993.34(1)：21-23.
[115]江武.消渴从肝论治刍议[J].中国中医药信息杂志.1996.28(11)：9-12.
[116]王智明,魏子孝.从肝论治消渴(糖尿病)的临床体会[J].中国中医基础医学杂志,2001.7(3)：65-66
[117]杨家景.糖尿病从肝论治[J].福建中医药.1997.35(6)：28-31.
[118]郭善军.逍遥降糖饮治疗糖尿病60例[J].北京中医学院学报.1991.30(3)：36-38.
[119]孟翠霞.滋水清肝饮加减治疗糖尿病38例[J].湖南中医学院学报.1995，(2)：18-20.
[120]杜丹,张文芳.刘天胤等.龙胆泻肝汤在治疗肥胖型消渴中的应用[J].中医药学报.1995.(4)：45-47.
[121]杨晓晖.四逆散加味治疗消渴病肝气郁结证[J].上海中医药杂志.1997.35(5)：11-13.
[122]刘素荣.刘桂芝.孙丰雷等.糖尿病从肝论治的理论探讨[J].山东中医药大学学报.1999(5)：340-341.
[123]倪海祥.刘刚，罗苏生等.从肝论治2型糖尿病的临床研究[J].中国中西医结合杂志，2000.20(8)：577-579
[124]鞠祥宗.韦淑萍.从肝论治糖尿病40例[J].陕西中医.2003.24(3)：201-202.
[125]周潮,李晓哲，单友琴等.降糖煎治疗II型糖尿病的临床与实验研究[J].光明中医杂志,1996,28(6)：51-54
[126]周劲刚.2型糖尿病从肝论治[J].四川中医.2012，30(4)：35-36
[127]徐志伟,敖海清,王文竹等.逍遥散对慢性心理应激损伤大鼠海马神经元的影响.中药理与临床,，2005,(21)1：5-8.
[128]徐志伟,敖海清,王文竹等.逍遥散对应激大鼠海马CA3区突触体结构可塑性的影响.中国行为医学科学杂志,2006,(15)2：102-106
[129]敖海清，徐志伟,苏俊芳等.柴胡疏肝散及逍遥散对慢性心理应激大鼠血清皮质酮及胃肠激素的影响.中药新药及临床药理,2007,(18)4：288-291.
[130]敖海清,徐志伟,富文俊等.逍遥散对慢性应激大鼠下丘脑-垂体-肾上腺轴调控作用机制的初步研究.中华行为医学与脑科学杂志.2010, (19)8：680-682.
[131]李光伟，潘孝仁,]Lillioja S等.检测人群胰岛素敏感性的一项新指数.中华内科杂志，1993，32(10)：656-660.
[132]Zimmet P,Alberti KG,Shaw J.Global and societalimplications of the diabetes epidemic [J].Nature,2001.414(6865):782-780.
[133]De Fronzo RA.Tobin J D, Andres R. Glucose clamp technique:a method for quantifying insulin secretion and resistance [J]. Am JJOURNAL OF PRACTICAL DIABETOLOGY Vol.7 NoPhys-iol 1979.237 (3):214-223.
[134]Matthews D R, Hosker J P.Rudenski A S.et al.Homeostasismodel assessment:insulin resist-ance and beta-cell function from fastingplasma glucose and insulin concentrations in man.Dia-betologia [J].1985.28(7):412-419.
[135]Ravipat IG,Aronow WS,Ahn C. et al.Association of hemoglobin Alc levelwith the severity of coronary artery disease in patients with diabetes mellitus[J].Am J Cardiol.2006,97(7):968-969.
[136]王喜,郑华丽.120例糖尿病患者血脂血糖的监测与分析[J].中国医药科学.2011，1(12)：111-112.
[137]McCann BS.Benjamin GA,W ilkinson CW. et al. Plasma lipid concentrationsduring episodic occupa- tional stress[J].Ann BehavMed,1999,21(2):103-110.
[138]Gottdiener JS,KopWJ.HausnerE, eta.l Effects ofmentalstress on flow-mediated brachial arterial dilation and influence ofbehavioral factors and hypercho-lesterolemia in subjects without cardio-vasculardisease[J].Am JCardiol 2003.92(6):687-691.
[139]Bacon SL,Ring C.Lip GY,et al.Increases in lipids and immune cells in response to exercise andmental stress in patientswith suspected coronary arterydisease:effects of adjustment for shifts in plasma volume[J].Biol Psychol.2004.65(3):237-250.
[140]Bachen EA,MuldoonMF,MatthewsKA.eta.l Effects ofhemoconcentration andsympathetic activation on serum lipid responses to briefmental stress[J]. PsychosomMed.2002.64(4):587-594.
[141]Chandak GR, Janipalli CS, Bhaskar Set al. Common variants in the TCF7L2 gene are strongly associated with type 2 diabetes mellitus in the Indian population [J]. Diabetologia,2007,50(1):63-67.
[142]王旭东.丁选平.2型糖尿病相关基因研究进展.中国优生与遗传杂志，2006，14(1)：4-5.
[143]罗敏.2型糖尿病相关基因的研究现状及展望.国外医学内分泌学分册，2002.22(3)：141-145.
[144]Mangion J. Rahman N. Mansour S. et al. A genefor lymphedema- distichiasis maps to 16q24.3 [J]. Am JHum Genet,1999,65:427-432.
[145]Cerderberg A, Gronning LM, Ahren Bet al. FOXC2 is a winged helix gene that counteracts obesity, hypertriglyceridemia, and diet-induced insulin resistance[J].Cell.2001.106:563-573.
[146]Jasonk Kim,Hyo-Jeong Kim et al.Adipocyte-specific overexpression of FOXC2 prevents diet-induced increases in intramuscular fatty acyl coa and insulin resistance[J].Diabetes,2005.54:1657-1663.
[147]YangX, SmithU.Decreased adipose tissue FOXC2 expression in insulin resistance-impaired precursor cell commitment to bothwhite andbrown adipose differentiation [C]. ADA 63rd Scientific Sessions,2003:381.
[148]DahleMK,Gronning LM, Cederberg A, et a.l Mechanisms of FOXC2and FOXD1-mediated regulation of the RI alpha subunit of campdependentprotein kinase include release of transcript-tional repressionand activation by protein kinaseB alpha and cAMP[J]. JBiolChem.2002,277(25): 22902-22908.
[149]Di Gregorio GB, Westergren R. Enerback S. et a.l Expression ofFOXC2 in adipose and muscle and its association with whole bodyinsulin sensitivity[J].Am J Physiol EndocrinolMetab.2004,287. (4):E709-E803.
[150]Kim JK.Kim HJ, Park S, et a.l Adipocyte-specific overexpression ofFOXC2 prevents diet-induce increases in intramuscular fatty acylCoA and insulin resistance[J].Diabetes,2005,54:1657-1673.
[151]YuC,ChenGW. ZhangD, eta.lMechanism bywhich fatty acids inhibit insulin activation of insulin receptor substrate-1(IRS-1)-associated phosphatidylinositol 3-kinase activity in muscle[J], J BiolChem, 2002.277:50230-50236.
[152]De Souza CT. Araujo EP, Stoppiglia LF. et al. Inhibition ofUCP2 expression reverses dietinduced diabetes mellitus byeffects on both insulin secretion and action[J]. FASEB J.2007,21(4):1153-1163.
[153]Pi J. Bai Y. Daniel KW. et al. Persistent oxidative stress dueto absence of uncoupling protein 2 associated with impairedpancreatic beta-cell function[J].Endocrinology.2009,150 (7):3040-3048.
[154]Affourtit C, Jastroch M. Brand MD. Uncoupling protein-2attenuates glucose-stimulated insulin secretion in INS-1Einsulinoma cells by lowering mitochondrial reactive oxygenspecies[J].Free Radic Biol Med,2011.50(5):609-616.
[155]Divakaruni AS, Brand MD.The regulation and physiology ofmitochondrial proton leak [J]. Physiology (Bethesda).2011.26(3):192-205.
[156]Leloup C, Tourrel-Cuzin C, Magnan C. et al. Mitochondrialreactive oxygen species are obligatory signals for glucose-induced insulin secretion[J]. Diabetes.2009.58 (3):673-681.
[157]Lablanche S.Cottet-Rousselle C.Lamarche Fet al.Protectionof pancreatic INS-1 beta-cells from glucose and fructose-induced cell death by inhibiting mitochondrial permeabilitytransition with cyclosporin A or metformin[J]. Cell DeathDis.2011.2(1):134-145.
[158]Pi J. Collins S. Reactive oxygen species and uncouplingprotein 2 in pancreatic beta-cell functional]. Diabetes ObesMetab.2010.12(Suppl 2):141-148.
[159]Harmon JS. Stein R. Robertson RP. Oxidative stress-mediated, post-translational loss of MafA protein as acontributing mechanism to loss of insulin gene expression inglucotoxic beta cells[J]. J Biol Chem,2005,280(12):11107-11113.
[160]Robertson RP. Chronic oxidative stress as a centralmechanism for glucose toxicity in pancreatic islet beta cells indiabetes[J]. J Biol Chem,2004,279(41):42351-42354.
[161]Pi J, Bai Y, Daniel KW, et al. Persistent oxidative stress dueto absence of uncoupling protein 2 associated with impairedpancreatic beta-cell function [J]. Endocrinology,2009,150(7):3040-3048.
[162]Affourtit C, Jastroch M, Brand MD. Uncoupling protein-2attenuates glucose-stimulated insulin secretion in INS-lEinsulinoma cells by lowering mitochondrial reactive oxygenspecies [J]. Free Radic Biol Med,2011,50(5):609-616.
[163]BraissantO,FouflleF,Scotto C et al. Differential expression of peroxisome proliferator--activated receptors (PPARs):tissue distribution of PPAR-alpha,-beta, and-gamma in the adult rat [J]. Endocrinology,1996,137(1):354-366.
[164]Akiyama T E, Nicol CJ, Fievet C et al. Peroxisome proliferator-activated receptor-alpha regulates lipid homeostasis, but is not associated with obesity studies with congenic mouse lines [J]. J Bi1 Chem, 2001,276(42):39088-39093.
[165]Li P,Zhu Z,Lu Y,et al. Metabolic and cellular plasticity in white adipose tissue O:role of peroxisome proliferator-activated receptor [J].Am J Physiol Endocrinol Metab,2005.289(4):E617-E626.
[166]Anandharajan R, Pathmanathan K, Shankernarayanan N P, et al. Upregulation of Glut-4 and PPARgamma by an isoflavone from Pterocarpus marsupium on L6 myotubes:a possible mechanism of action [J]. Ethnopharmacol,2005,97(2):253-260.
[167]Hegele RA.Monogenic forms of insulin resistance:apertures that expose the common metabolicsyndrome [J]. Trends Endocrinol Metab,2003,14(8):371-377.
[168]Enomoto N.Takei Y. Hirose M, et al. Prevention of ethanol induced liver injury in rats by an a gonist of peroxisome proliferators-activated receptor-gamma pioglitazone [J].Pharmacol ExpT--her.2003, 306(3):846-854.
[169]Barens PJ, Adock IM. Anti-inflammatory actions of steroids; molecularmechansims[J].Trends Pharmacol Sci,1993.14:436-441.
[170]Delaunay F. KhanA, Cintr A, etal. Pancreatic beta cells are important targets for the diabetogenic effects of glucocorticoids [J]. J Clin Invest,1997,100:2094-2098.
[171]Ling ZC,KhanA.DelaunyF,et al. Increased glucocorticoid sensitivity in islet beta-cells:effectson glucose-6-phosephatase,glucose cycling and insulin release [J]. Diabetologia,1998.41:634-639.
[172]Itoi K, Jiang YQ, Iwasaki Y. et al. Regulatory mechanism ofcorticotrophin-releasing hormone and va--sopressin gene expression inthe hypothalamus[J]. Neuroendocrinol,2004,16(4):348-355.
[173]ChanO.Inouye K.AkiravEM,et al.Hyperglycemia does notincrease basal hypothalamo-pituitary-adrenal activity in diabetes butit does impair the HPAresponse to insulin-induced hypoglycemia[J].Endocrinology.2005.146(3):1382-1390.
[174]崔俊芳，张如意.2型糖尿病患者的血浆皮质醇水平的变化及与代谢指标的相关分析[J].宁夏医学杂志,2010.32(6)：535-537.
[175]Evans SB.Wilkinson CW.Gronbeck P,et al.Inactivation of the DMHselectively inhibits the ACTH and corticosterone responses tohypoglycemia[J].Am J Physiol Regul Integr Comp Physiol,2004, 286(1):123-128.
[176]NyirendaMJ, LindsayRS.KenyonCJ.etal. Glucocorticoidexposure inlate gestation permanently programsrathepatic phosphoenolpyruvatecarboxy kinaseandglucocorticoidreceptorexpressionandcauses glucosein tolerance adultoffspring [J].JClinlnvest,1998.101:2174-2181.
[177]李芳萍,张四清,王斐等.2型糖尿病和非酒精性脂肪肝病的胰岛素抵抗研究[J].新医学,2009.40(7)：427-30.
[178]刘淑娥,肖丹，肝源性糖尿病临床分析[J].中日友好医院学报,2003,17(3)：153-154.
[179]Myers G M Jr, Sun X J,et al. The IRS-1 signaling system. TIBS,1994,19(7):289-293
[180]唐伟,朱剑,武晓泓等.胰岛素信号转导及其调节机制[J].实用糖尿病杂志,2005,1(3)：43-49.
[181]周云枫,吴勇,欧阳静萍.黄芪多糖对2型糖尿病大鼠肾组织胰岛素信号转导的影响[J].武汉大学学报,2005,26(2)：139-143.
[182]Aspinwall CA, Qian WJ, Roper MG, et al. Roles of insulin receptorSubstrate-1. phosphatidylinosi--tol 3-kinase, and release of intracellular Ca2+ stores in insulin-stimulated insulin secretion inβ-cells[J]. J BiolChem.2000,275(29):22331-22338.
[183]张妍，袁莉.p细胞胰岛素受体信号系统与p细胞功能[J].国外医学内分泌学分册，2005,25(4)：259-261.
[184]裴海成.高浓度游离脂肪酸对胰岛素受体后信号转导的下降调节[J].第二军医大学学报，2003,24(5)：569-573.
[185]Yuan L.Ziegter R.Hamann A.Meffomdn modulates insulin post-receptorsignalingtrans- duction in chronically insulin-treated HepG2 cells[J]. Acta Pharmacol Sin.2003,24(1):55-60.
[186叶‘华.2型糖尿病患者及其一级亲属和非糖尿病者骨骼肌的胰岛索信号传导[J].中国糖尿病杂志，2003，11(2)：151-152.
[187]彭定琼，高妍,陈宇等.OLETF大鼠肝脏、肌肉、脂肪组织中胰岛素受体底物1的蛋白表达[J].中华内分泌代谢杂志.001，17(5)：288-289.
[188]张中成，刘志诚.2型糖尿病胰岛素受体后水平研究现状[J].医学综述，2002，8(3)：166-167.